System for releasing and isolating nucleic acids

An instrument that can hold one or more sample processsing vessels, maintain the sample processing vessels at a constant temperature, shake the sample processing vessels and separate magnetic particles by means of magnetic force. This system greatly simplifies the isolation of nucleic acids.

The object of the invention is a system for the release and isolation of 
nucleic acids and a procedure for using this system. 
Detection procedures based on the determination of nucleic acids in a 
sample have increased in significance recently. This is due, among other 
things, to the high sensitivity of detection that these procedures can 
achieve. In terms of sensitivity, nucleic acid detection procedures are 
basically superior to antigen detection procedures. Although antigens are 
often relatively accessible in a sample, numerous steps are usually 
required to make nucleic acids accessible, especially when detecting 
organisms. In addition, nucleic acids are usually present in very low 
concentrations. Purification procedures for isolating nucleic acids from 
samples containing cells are known in particular, although they require a 
great deal of time and effort. 
The sensitivity of the sample enrichment and pretreatment systems for 
nucleic acids currently on the market is often insufficient. In addition, 
automated sample pretreatment systems are not sufficiently safe from 
contamination to enable amplification, e.g. using the PCR. Another 
disadvantage of the automated sample pretreatment systems currently 
available is that they require the use of organic solvents (phenol and/or 
chloroform alcohol mixtures) to extract the nucleic acids. 
The procedures in use today that immobilize nucleic acids basically use two 
principles to isolate nucleic acids. One principle calls for liquid 
samples containing nucleic acids to be aspirated through a solid phase 
which retains the nucleic acids. This step is preceded by a lysis step 
performed in a separate container. The nucleic acids are then dissolved 
from the solid matrix by aspirating an elution fluid through the matrix. 
The elation solution containing the nucleic acids is aspirated into a 
container for the next steps. It has been demonstrated, however, that the 
purity of the devices in use today does not meet the requirements for a 
subsequent amplification reaction, such as the PCR. 
According to the second principle of nucleic acid isolation, the nucleic 
acids are removed by way of precipitation and then separated in a 
centrifuge. This procedure cannot be performed in a "batch" mode, however. 
Rather, it first requires that a solution containing cells be treated with 
lysing agents in a reaction vessel. The reaction mixture is then 
transferred by pipette from the container to a centrifugation tube. This 
tube contains an insert to which the released nucleic acids can adsorb, 
while the remaining fluid can flow to the bottom of the tube during 
centrifugation. The insert is treated one or more times with a fluid to 
wash the absorbed nucleic acids. For this step, the insert is transferred 
to a second centrifugation tube so that residues from the sample fluid do 
not reenter the insert. In the final step, the insert is placed in yet 
another container. An elution solution is then centrifuged through the 
insert to transfer the nucleic acids to another vessel that contains a 
solution that is capable of being processed further. This procedure is 
very susceptible to contamination, however, and requires transferring 
solutions between numerous reaction vessels. 
The task of this invention was to provide a system for which the 
disadvantages of the state of the art are eliminated either completely or 
at least partially. In particular, this system can be used to absorb and 
desorb nucleic acids to a solid phase matrix without requiring a 
centrifuge for these steps. 
A main feature of the invention is a receptacle for a sample processing 
vessel that can be kept at a constant temperature and set into motion in 
order to thoroughly mix the substances contained in the sample processing 
vessel. In addition, this receptacle is connected to a vacuum-generating 
system (e.g. a hose pump or a piston pump). The receptacle also makes it 
possible to separate magnetic particles within the sample processing 
vessel. Important advantages of the invention are the protection it 
provides against contamination (between samples and between the system and 
the environment), and its potential to hold enough sample processing 
vessels to ensure cost effectiveness. 
This invention also provides a procedure for releasing and isolating or 
detecting nucleic acids from biological compartments of a sample with the 
following steps: 
The sample is incubated in a sample processing vessel with magnetic 
particles that can bind with the biological compartments while the sample 
processing vessel is shaken, 
A magnet is positioned near the sample processing vessel in order to hold 
the magnetic particles against the wall of the vessel, 
The remaining fluid is removed from the sample processing vessel, 
The magnetic particles are resuspended in a second fluid by 
a) removing the magnet away from the sample processing vessel so that the 
magnetic particles are no longer held against the wall of the vessel, 
while 
b) shaking the sample processing vessel, 
The biological compartments are warmed and lysed, 
The lysis mixture is cooled under conditions that make it possible to 
immobilize or hybridize the nucleic acids to be isolated or detected. 
This invention also provides a procedure for the release and isolation of 
nucleic acids from a suspension of biological compartments using magnetic 
particles with the following steps: 
A sample is incubated in a sample processing vessel with magnetic particles 
in order to lyse the biological compartments, 
The lysis mixture is cooled and the nucleic acids to be isolated or 
detected are immobilized on the magnetic particles, 
The state of immobilization is eliminated and the nucleic acids to be 
isolated and purified are transferred to a vessel from which they can be 
pipetted.

Nucleic acids" as described in context with this invention refer to nucleic 
acids that are present in biological compartments. "Biological 
compartments" refer in particular to viruses or cells of bacterial origin 
for instance. In an especially preferred construction, the cells are 
basically present in a format in which they are basically separate from 
each other. In principle, however, this invention can also process 
multicellular compartments. These compartments and their nucleic acids are 
contained in a sample at the beginning of the procedure described by this 
invention. This sample is preferably a suspension of biological 
compartments in a fluid. These fluids can be obtained from bodily fluids 
such as blood, saliva or urine, for instance. 
"Release of nucleic acids" as described in context with this invention 
refers to the discharge of nucleic acids from the biological compartments. 
Any means can be used to discharge the nucleic acids from the biological 
compartments. Preferably, the wall separating the biological compartments 
from the fluid is destroyed. This can be accomplished, for instance, by 
treating the compartments with cell wall destroying agents such as 
proteinase K. "Isolation of nucleic acids" as described in context with 
this invention refers to the separation of nucleic acids from other 
components in the sample. "Other components in the sample" can include the 
walls of the biological compartments, their degradation products, other 
contents of the biological compartments and components of the fluid that 
surrounds the biological compartments in the sample. These components can 
include proteins or enzyme inhibitors, especially enzymes that degrade 
nucleic acids, such as Dnase or RNase. In this sense, isolation can also 
refer to a method for purify nucleic acids. This isolation step can be 
specific or non-specific for other nucleic acids contained in the sample. 
"Detection of nucleic acids" as described in context with this invention 
refers to a procedure in which the presence or quantity of nucleic acids 
is determined. These procedures can be performed quantitatively or 
qualitatively. The quantitative detection procedure usually requires that 
a comparison test be performed with a sample that contains a known 
quantity of the nucleic acids to be detected. The detection can be 
sequence-specific or sequence non-specific. To make the detection 
specific, one usually uses "probes" that are characterized by the fact 
that they have a nucleobase sequence that is more or less characteristic 
for the nucleic acids in the sample. If the goal is to perform specific 
detection of nucleic acids, a probe is used that contains a base sequence 
that is complementary to the base sequence of the nucleic acids to be 
detected, but not, however, to other nucleic acids in the sample. Probes 
can be molecules that contain a group that can be detected either directly 
or indirectly. Groups that can be detected directly include radioactive 
(.sup.32 P), colored or fluorescent groups or metal atoms. Groups that can 
be detected indirectly include, for instance, compounds with an 
immunological or enzymatic effect such as antibodies, antigens, haptens, 
enzymes or enzymatically active sub-enzymes. These groups are detected in 
a subsequent reaction or sequence of reactions. Especially preferred are 
haptens, such as digoxigenin or biotin. Hapten-labelled probes of this 
nature can then be detected easily in a reaction with a labelled antibody 
against the hapten. 
DESCRIPTION OF THE SYSTEM 
The object of the invention is a system for the release and/or isolation of 
nucleic acids from a suspension of biological compartments comprising the 
following components: 
a receptacle (10) for one or more sample processing vessels (A), 
a thermostat unit (20) to maintain the sample processing vessels (A) and 
the fluids they contain at a constant temperature, 
a mechanical shaker (30) to shake the sample processing vessels (A), 
a separation device (40) to separate magnetic particles from fluid using 
magnetic force and deposit them on a wall of each sample processing vessel 
(A), and 
a pump unit (50) to remove fluid from the sample processing vessel (A). 
The surface of the system is easy to clean and protects the operator from 
burns (e.g. by virtue of a plastic housing). 
Drawings of a system comprising the units described by this invention are 
shown in FIGS. 1 and 2. 
The system has a receptacle (10) for holding the sample processing vessels. 
This receptacle contains numerous recesses (12) or "cavities" into which 
the sample vessels are placed. The cavities are preferably arranged in a 
linear format or in subgroups that are arranged in a linear format yet are 
at right angles to each other. The distance between the cavities is 
preferably the same distance that separates the wells of a microtiter 
plate and, more preferably, twice this distance. The cavities are designed 
to hold the sample vessels. 
The sample processing vessel (A) can basically have any shape or design. 
For instance, these sample processing vessels can be the wells of a 
microtiter plate (e.g. a 96-well format). The vessels are preferably 
cylindrical, however, with an opening at the top to receive fluid and, 
most preferably, an opening at the bottom from which fluid can exit (A11). 
A sample processing vessel with this design can be used to reduce 
contamination while processing samples containing nucleic acids. These 
vessels are usually made of a plastic such as polypropylene. 
In an especially preferred construction of this invention, vessels can be 
used that are described in the German design patent applications numbered 
29505652.5 and 29505707.6. 
The fluid in the sample processing vessels is kept at a constant 
temperature in the system described by this invention by means of a 
thermostat unit (20). This unit, which basically consists of components 
common to thermostats, is preferably integrated--at least partially--in 
the receptacle (10) in which the sample processing vessels can be placed. 
The receptacle (10) especially includes a metal block with good thermal 
conductivity. The shape of this metal block is designed to fit with the 
outer contour of the sample processing vessels in such a way that the 
walls of the sample processing vessels placed in the receptacle fit within 
the metal block as snugly as possible, so that heat can be transferred 
efficiently. The temperature in the block is increased or decreased, 
depending on the reaction to be performed in the sample processing vessel. 
The temperature can range from 4.degree. C. to 95.degree. C. Ideally, 
numerous electrical heating elements located as closely as possible to the 
sample processing vessel are used to increase the temperature. The fluid 
in the sample processing vessel is cooled by means of peltier elements 
that are also located as close to the sample processing vessels as 
possible. The temperature is regulated preferably by means of a fuzzy 
regulator or a PIDT regulator. A sufficient number of heat sensors are 
situated in proper locations within the thermostat unit. 
Another construction of the thermostat unit (20) includes a block through 
which fluid flows. Fluids--usually water or aqueous saline solutions--are 
used as the incubation medium. Preferably, the incubation fluid is fed 
directly into openings (21) in the block (10) by means of a circulating 
pump via flexible tubes connected to a heating or cooling system. The 
volume of the incubation reservoir (represented in FIG. 1 with "heating 
unit" and "cooling unit") outside the DNA module is much larger than the 
dead volume of the DNA module in order to minimize this disturbance 
variable when the 2-way valve is switched over. The heating and cooling 
units maintain the fluid at a constant temperature before the process 
begins and can be programmed to operate at certain intervals if necessary. 
A regulator controls the valve and heating and cooling units, and, coupled 
with the appropriate volume flow achieved by the circulating pump, 
maintains the heating and cooling rates desired. The "thermostat unit" 
(20) in this construction refers to a combination of a block, circulating 
pump, heating unit, cooling unit and 2-way valve through which fluid 
flows. The "DNA module" refers to the device that includes the receptacle 
for the sample processing vessel, the shaking device and the separation 
device. 
The system enables one to work with magnetic particles (beads). The term 
"magnetic particles" refers to particles that can be transported in a 
certain direction by means of magnetic force. These particles can be made 
of ferromagnetic or superparamagnetic materials, for instance. Especially 
preferred in the context of this invention are ferromagnetic materials. 
Particles are solid materials with a small diameter. Within the context of 
this invention, particles with an average particle size of more than 2.8 
.mu.m but less than 200 .mu.m are especially suitable. Most preferably 
they have an average particle size of between 10 and 15 .mu.m. The 
distribution of particle size is preferably homogenous. The surface of 
these particles is modified in such a way that they can bind with the 
biological compartments. Magnetic particles that are suitable for this 
application are the known and commercially available latex magnetic 
particles to which antibodies can be bound, for instance. Antibodies 
targeted against surface antigens of the biological compartments are used 
in particular to bind the biological compartments to the magnetic 
particles. Magnetic particles of this nature are also commercially 
available. 
Glass-magnetic pigments with surfaces to which nucleic acids can bind can 
also be used. Glass-magnetic pigments of this nature are known from the 
German patent application number 19537985.3. 
For the procedure described by this invention to be performed successfully, 
the magnetic particles must be bound to the inner wall of the sample 
processing vessel for certain reaction steps and then brought back into 
suspension in a subsequent step. In an especially preferred construction 
of this invention, a separation device (40) to which one or more permanent 
magnets or electromagnets are attached is moved towards the sample 
processing vessel in order to position the magnets. The removal of the 
magnet away from the sample processing container--which is necessary for 
separation to be performed efficiently--depends to a large extent on the 
strength of the magnetic field that the magnets can produce, the size of 
the magnetic particles, and on the ability of the magnetic particles to be 
magnetized. The nature of the subsequent processing steps (e.g. mechanical 
stressing of the magnets) also determines the strength of the magnetic 
field to be used. If a permanent magnet is used, it is moved from a 
position where it cannot separate the magnetic particles and into the 
vicinity of the vessel so that the magnetic particles are held against the 
vessel wall. If an electromagnetic is used, it is turned on and allowed to 
remain in the power ON state until the biological compartments held 
against the vessel wall are processed. "Positioning a magnet in the 
vicinity of the vessel" also refers to the case in which the vessel is 
brought into the vicinity of the magnet Ultimately, it refers merely to 
the motion of the magnet in relation to the vessel. The separation device 
(40) preferably has a magnet that can be moved towards the sample 
processing vessel along a predetermined path, e.g. on rails or, 
preferably, by moving the magnet on a circular track, e.g. along an axis 
that passes next to the sample vessel. The separation unit also comprises 
a motor that can drive the movement of the magnet towards and away from 
the sample processing vessel. 
In another construction, the separation unit (40) has a gear rack that can 
be driven in linear fashion by means of a d.c. motor or a stepping motor. 
A device that holds the magnets--permanent magnets in this case--is 
positioned at a right angle to the gear rack. Light barriers can be used 
to detect the end positions of the movement of the magnets. In the 
preferred construction, one magnet is assigned to each sample processing 
vessel, the front face of which is moved into position next to the vessel. 
Two magnets can also be moved into position next to each sample processing 
vessel, however. Preferably, one magnet is moved into position next to 2 
vessels, so that only n+1 magnets are required for n vessels. 
For the separation step, the magnets must be moved as close to the sample 
processing vessels as possible in order to achieve a high rate of 
separation of the magnetic particles. It is also important that the 
distance traveled by the magnets between the positions be sufficiently 
long. Depending on the material and geometry of the magnet used, the 
magnets must be up to 40 mm away from the sample processing vessel to 
prevent unintentional separation of the magnetic particles in the sample 
processing vessel. For the instrument to function properly, it is 
important that the magnetic forces only affect the particles in the sample 
processing vessel, if this is so desired. To achieve this effect, the 
inactive position of the magnet must be far enough away from the vessel 
that the magnetic field has no effect on the movement of the particles. 
This effect can also be achieved in such a way that the distance between 
the magnet and the sample processing vessel is not changed. The magnetic 
fields can simply be interrupted by a .mu.-metal that is moved into a 
location between the vessel and the magnet. 
In another possible construction, the magnets are arranged on a rotatable 
shaft driven by a d.c. motor. This construction enables the magnets to be 
moved along a circular path. The magnets themselves can be arranged on 
this shaft in any order that moves them towards or away from the sample 
processing vessels. In addition, a drive mechanism is preferred in which 
one shaft with a number of magnets is situated on each opposing side of 
the system. The shafts are driven by a motor by means of a gear rack. With 
this arrangement, every two magnets move in a synchronous motion towards 
the front face of the sample processing vessel. 
Magnets for the invention described here preferably have a mass of between 
0.5 and 5 g, and, especially preferred, between 1 and 4 g. The outer 
dimensions of the prototype are 10 mm.times.10 mm.times.3 mm. Materials 
that have been proven to be suitable for a permanent magnet are rare earth 
materials (e.g. NeFeB, VACODYM 370 HR) with an optimal BH maximum at the 
smallest dimensions. For the separation step to proceed efficiently, it is 
advantageous to design the gradients of the magnetic field to be 
especially pronounced. For this reason, the magnets should be located as 
close to the vessel as possible. The sample processing vessels are 
preferably made of materials that weaken the magnetic field as little as 
possible, such as polypropylene. 
Tubes (51) that are connected to a vacuum-generating system are attached to 
the underside of the cavity on the unit that holds the sample processing 
vessels. One tube is assigned to each cavity. Since there are openings in 
the bottom of the sample processing vessels, a vacuum can be created to 
aspirate the contents of the sample processing vessels and deposit them in 
the waste container. In the construction shown in FIG. 2, each cavity has 
a seal that prevents air from being introduced into the space between the 
sample processing vessel and the inlet (14) when the waste material is 
aspirated. 
The vacuum system basically comprises a piston pump (50) that is connected 
with the cavities via a tubing system. A waste container into which the 
fluid is deposited is located between the piston pump and the cavities. 
The fluid that is aspirated out of the sample processing vessels is waste. 
In addition, a valve is situated between every sample processing vessel 
and the waste container. This valve enables the vacuum to be switched to 
each cavity when the system is permanently evacuated from the pump, past 
the waste container, and up to the valve. With the construction provided 
by the invention, the sample processing vessels can be aspirated in 
parallel and sequentially. 
In another construction, piston pumps and valves are replaced with a flow 
inducer. The waste container is not evacuated permanently in this case, 
rather, it is situated in the fluid stream behind the cavities and the 
flow inducer. With this arrangement, the cavities can only be aspirated in 
parallel. Numerous flow inducers may be used as well, which then serve a 
certain number of cavities and enable work to be performed in a partially 
sequential fashion. 
The sample processing vessels are preferably moved in a horizontal 
direction. In an especially preferred construction, the receptacle 
(10)--which contains recesses (12) that hold one sample vessel each--is 
moved, so that all sample vessels in the system are shaken. Vibration 
absorbers (11) serve to reduce the amount of movement transferred to the 
rest of the instrument. This invention preferably uses a mechanical shaker 
(30) that moves the sample processing vessels (A). This unit can basically 
be any mechanical device that is suitable for mixing fluids in a vessel. A 
preferred example of such a unit is described below. 
A stepping motor with an eccentric cam and an equalizing weight situated on 
a fixed framework (1) move the receptacle--which is placed on vibration 
absorbers on this framework--in a circular, eccentric path with a fixed 
amplitude and a variable frequency. The preferred amplitude A is 
.ltoreq.1.5 mm, and the preferred frequency (f) is greater than 1 Hz and 
less than 50 Hz. The mixing and resuspension step lasts between 5 and 30 
s, depending on the physical characteristics of the sample material. The 
amplitude can be varied by replacing the eccentric cam. 
Combining the system provided by this invention with an automated pipetting 
system is not a logical step, because this would require that the sample 
vessels be placed in a definite position before and during the pipetting 
steps. If the sample vessels are not placed in a definite position, they 
are located in a different position after the shaking step. If the 
position of the vessels during the pipetting step is not specified 
exactly, it may not be possible to perform the pipetting procedure 
correctly. For this reason, the instrument ensures that the vessel is 
located in a defined "home" position after the shaking procedure, from 
where a pipetting step or other processes can be performed. 
It is advantageous to use a stepping motor instead of a d.c. motor in order 
to ensure a defined home position. In a preferred construction, the home 
position is detected with a light barrier. 
The instrument may also be designed to be non-invasive, as described below. 
However, these alternatives are more complex in design (Versions 1 and 2), 
or they utilize mixing steps (3) that take a longer time to complete: 
1. A combination of one, two or three linear drives for the receptacle on 
the plane or in a space (X, Y and Z-axis) to create Lissajous curves, for 
instance. 
2. Wobbling by tilting the framework (1) at a certain angle and placing the 
receptacle (10) at the opposite end. 
3. Magnetic stirring mechanism 
4. Swirling or tapping the DNA module. 
The system components are coupled in such a way as to be functional, e.g. 
via integration of the magnets in the unit (10). The system components are 
also coupled chronologically. For instance, the units are operated in the 
sequence required for the application desired, e.g. via a computer program 
or by the operator initiating the steps individually. 
Description of the Procedure 
Version A 
In the initial step, the sample is incubated in a sample processing vessel 
with magnetic particles (beads) that can bind with the biological 
compartments while the sample processing vessel is shaken. 
The incubation of the samples with the magnetic particles can take place in 
any fashion. It is necessary for the sample and the magnetic particles to 
be placed in the sample processing vessel. Neither the method by which the 
sample and magnetic particles are placed in the vessel nor the sequence in 
which this takes place is especially significant for the procedure 
provided by this invention. Preferably, however, the magnetic particles 
are pipetted into the sample processing vessel in a suspension with a 
known concentration of magnetic particles. The sample is pipetted into the 
sample processing vessel either before or after the suspension of magnetic 
particles. 
The mixture is incubated under appropriate conditions until a sufficient 
quantity of biological compartments are bound to the magnetic particles, 
usually between 1 minute and 10 minutes. The sample processing vessel is 
preferably closed in proper fashion, e.g. with a cap and/or a valve. 
An important feature of the invention is the fact that the mixture in the 
sample processing vessel is shaken during incubation. The mixture can be 
shaken in intervals, or it can be shaking during the entire incubation 
period or only during certain periods. The mixture is shaken in order to 
sufficiently mix the biological compartments and the magnetic particles in 
the fluid, and especially to suspend or resuspend the beads and accelerate 
diffusion. This reduces the amount of time required to bind the biological 
compartments to the magnetic particles. 
After the incubation step and after the compartments are bound to the 
magnetic particles, the biological compartments are removed from the 
surrounding fluid in the sample. An appropriate method of accomplishing 
this is to collect the magnetic particles and bound biological 
compartments by positioning a magnet near the sample processing vessel. 
This holds the magnetic particles with the biological compartments against 
the vessel wall, as is preferable. The beads are therefore usually 
separated on the inside wall of the sample processing vessel or a part 
thereof that is located below the surface of the sample fluid. 
The fluid surrounding the biological compartments is then removed from the 
sample processing vessel. This is performed under conditions in which the 
magnetic particles remain against the vessel wall. The method used to 
remove the sample depends on the type of sample processing vessel used. 
The fluid can be removed via pipette, for instance. In a preferred 
construction, however, in which the sample processing vessel has an 
opening at the bottom from which the fluid can exit, the fluid is simply 
aspirated through this opening. This fluid removal method minimizes the 
mechanical stress placed on the magnetic particles and thereby prevents 
them from being removed from the vessel wall. 
An especially important step is the resuspension of the magnetic particles 
that remain on the vessel wall in a second fluid that is added. To 
accomplish this, the magnet is moved away from the vessel so that it no 
longer holds the magnetic particles against the vessel wall. As described 
above, it is also possible to move the vessel away from the magnet. With 
regard for the invention described here, it has been demonstrated that 
simply removing the magnet is not enough to resuspend the magnetic 
particles in the solution if the vessel is not also shaken, and preferably 
simultaneously. This shaking motion is performed by the mechanical shaker 
(30). It causes the magnetic particles to be distributed evenly within the 
second fluid. This second fluid can be added to the sample processing 
vessel, e.g. via pipette, before or after the magnet is removed. 
The procedure provided by this invention can also be used to further purify 
biological compartments. To accomplish this, a suspension of magnetic 
particles that bind with the biological compartments is positioned in a 
sample processing vessel in relation to a magnet in such a way that the 
magnetic particles with the biological compartments are held against the 
vessel wall. The fluid that contains the biological compartments is then 
removed from the vessel and the magnetic particles are resuspended in a 
second fluid--a wash fluid in this case--by moving the magnet away from 
the vessel so that the magnetic particles are no longer held against the 
vessel wall, while the vessel is shaken. This wash step can be repeated as 
needed until the biological compartments have reached a sufficient level 
of purity. 
Another step of the procedure provided by this invention is the subsequent 
disintegration (lysis) of the biological compartments. Procedures for 
lysing biological compartments are known by the expert, as are the 
specific conditions for certain types of compartments, e.g. cells. To lyse 
bacteria, for instance, a mixture of proteinase K is added to the 
biological compartments and incubated for the amount of time necessary to 
lyse or partially or completely decompose the cell walls and release the 
nucleic acids contained in the biological compartments. This procedure is 
preferably performed at temperatures above room temperature, and more 
preferably, at temperatures between 70 and 95.degree. C. The mixture 
created when the cells are lysed is also referred to as the lysis mixture 
below. The incubation period is preferably from 5 to 20 minutes, and more 
preferably, from 10 to 15 minutes. If the cells are lysed at room 
temperature or a temperature slightly above room temperature, it is 
especially preferable to then warm the lysis mixture to higher 
temperatures such as 70.degree. C. or, if the samples are potentially 
infectious, to 95.degree. C. The lysis can also be deactivated if it 
interferes with the subsequent steps. 
The lysis mixture is then cooled under conditions that depend on the 
purpose for which the procedure provided by this invention is performed. 
If the nucleic acids are isolated on a solid phase, conditions are 
selected under which the nucleic acids can bind to the solid phase. A 
suitable procedure for binding nucleic acids is the incubation of released 
nucleic acids with glass surfaces in the presence of chaotropic salts. A 
procedure of this nature is described in EP-A-0 389 063, for instance. In 
this procedure, the nucleic acids are bound non-specifically to the glass 
surface, while other components of the biological compartments and the 
lysis reagent are not bound to the glass surface, or are only bound 
insignificantly. The fluid that contains the remaining components is then 
preferably removed from the sample processing vessel, e.g. via aspiration, 
while the glass surface can remain in the sample processing vessel with 
the nucleic acids bound to it. In a preferred construction, a solid phase 
in the form of a glass fiber fleece is placed in the sample processing 
vessel and incubated with the mixture. In this procedure, the nucleic 
acids are immobilized on the glass fibers and can simply be removed from 
the sample processing vessel with the glass fiber fleece. 
If the nucleic acids will be detected after their release, they are 
hybridized with a probe. This probe, as described above, is a molecule 
that has a base sequence that is complementary to the nucleic acid to be 
detected or a part thereof. In a preferred case, this is an 
oligonucleotide labelled with a group to be detected. The reaction mixture 
is therefore cooled under conditions in which the nucleic acids to be 
detected hybridize with the nucleic acid probe. These temperatures are 
known by the expert. In another construction of the procedure for 
detection of nucleic acids, the nucleic acids to be detected hybridize 
with a nucleic acid probe bound to a solid phase. In this procedure, the 
probe can be used on any solid phase, such as microtiter plate cavities or 
the inside wall of the sample processing vessel, as long as it is 
separated only from the rest of the reaction mixture. Procedures for 
immobilizing nucleic acid probes, especially "capture" probes, are known 
by the expert, e.g. from EP-A-0 523 557. 
After the mixture is cooled, the nucleic acids to be isolated or detected 
are then separated from the surrounding fluid that may still contain 
remains of the lysis mixture and reagents used to bind the nucleic acids 
to a solid phase. Depending on the type of solid phase used, the solid 
phase can be filtered or removed from the sample processing vessel, or the 
fluid can be removed from the sample processing vessel by pipette. 
The bound nucleic acids can then be unbound from the solid phase, detected 
directly in common procedures for detecting nucleic acid sequences known 
by the expert, or labelled. 
Version B 
In the initial step, the sample is pipetted into a sample processing vessel 
along with lysing reagent and glass-magnetic particles (beads) that can 
bind with the nucleic acids contained in the biological compartments. The 
sample processing vessel is then closed and shaken. It is necessary for 
the sample, lysis reagent and glass-magnetic particles to be placed in the 
sample processing vessel. Neither the method by which they are placed in 
the vessel nor the sequence in which this takes place is especially 
significant for the procedure provided by this invention. Preferably, 
however, the glass-magnetic particles are pipetted into the sample 
processing vessel in a suspension with a known concentration of 
glass-magnetic particles. The sample is pipetted into the sample 
processing vessel either before or after the suspension of glass-magnetic 
particles. It is essential that the sample, glass-magnetic particles and 
lysis reagent are shaken until mixed thoroughly. 
Another step of the procedure provided by this invention is the subsequent 
disintegration (lysis) of the biological compartments. Procedures for 
lysing biological compartments are known by the expert, as are the 
specific conditions for certain types of compartnents, e.g. cells. To lyse 
bacteria, for instance, a mixture of proteinase K is added to the 
biological compartments and incubated for the amount of time necessary to 
lyse or partially or completely decompose the cell walls and release the 
nucleic acids contained in the biological compartments. This procedure is 
preferably performed at temperatures above room temperature, and more 
preferably, at temperatures between 70 and 95.degree. C. The mixture 
created when the cells are lysed is also called the lysis mixture below. 
The incubation period is preferably from 5 to 20 minutes, and more 
preferably, from 10 to 15 minutes. 
If the cells are lysed at room temperature or a temperature slightly above 
room temperature, it is especially preferable to then warm the lysis 
mixture to higher temperatures such as 70.degree. C. or, if the samples 
are potentially infectious, to 95.degree. C. The lysis can also be 
deactivated if it interferes with the subsequent steps. 
An important feature of the invention is the fact that the mixture in the 
sample processing vessel is shaken during incubation. The mixture can be 
shaken in intervals, or it can be shaking during the entire incubation 
period or only during certain periods. The mixture is shaken in order to 
sufficiently mix the biological compartments and the magnetic particles in 
the fluid, and especially to suspend or resuspend the beads and accelerate 
diffusion. This reduces the amount of time required to bind the biological 
compartments to the magnetic particles. 
The lysis mixture is then cooled under conditions that depend on the 
purpose for which the procedure provided by this invention is performed. 
The nucleic acids released from the biological compartment should now bind 
non-specifically to the surface of the glass-magnetic particles. To 
improve the binding characteristics, i-propanol or ethanol--depending on 
the type of biological compartment involved--is added to the lysis mixture 
after lysis. The sample processing vessel is then shaken to mix the 
mixture further. The nucleic acids are now bound non-specifically to the 
surface of the solid phase. Other components of the biological compartment 
and lysis reagents do not adsorb to the glass surface, or only 
insignficant components adsorb to the glass surface. 
After the solid phase binding step is complete, the magnetic fields are 
activated in order to deposit the glass-magnetic pigment with the bound 
nucleic adds on the inside surface of the sample processing vessel. The 
remaining fluid is then removed from the vessel. In a preferred 
construction, however, in which the sample processing vessel has an 
opening at the bottom from which the fluid can exit, the fluid is simply 
aspirated through this opening. This fluid removal method minimizes the 
mechanical stress placed on the magnetic particles and thereby prevents 
them from being removed from the vessel wall. 
In a subsequent step, the glass-magnetic particles are resuspended in a 
wash fluid. To accomplish this, the magnet is moved away from the vessel 
so that it no longer holds the magnetic particles against the vessel wall. 
As described above, it is also possible to move the vessel away from the 
magnet. With regard for the invention described here, it has been 
demonstrated that simply removing the magnet is not enough to resuspend 
the magnetic particles in the solution if the vessel is not also shaken, 
and preferably simultaneously. This shaking motion is performed by the 
mechanical shaker (30) and causes the magnetic particles to be distributed 
evenly within the second fluid. This second fluid can be added to the 
sample processing vessel, e.g. via pipette, before or after the magnet is 
removed. This wash step can be repeated as needed until the biological 
compartments have reached a sufficient level of purity. 
The bound nucleic acids can then be unbound from the solid phase, detected 
directly in common procedures for detecting nucleic acid sequences known 
by the expert, or labelled. 
The procedure provided by this invention is therefore based on a 
combination of steps that make use of a receptacle (10) for holding one or 
more sample processing vessels, a thermostat unit (20) to maintain the 
sample processing vessels and the fluid in them at a constant temperature, 
a mechanical shaker (3) to shake the sample processing vessels, and a unit 
(40) for separating and depositing the magnetic particles on a wall of 
each sample processing vessel using magnetic force. Surprisingly, these 
steps and units can be combined into a single reaction block. A reaction 
block in this case refers to a device that comprises all or some of the 
units 10, 20, 30 and 40 coupled in coordinated fashion. This invention 
enables a process to be performed in one instrument that used to require 
numerous manual preparation steps. The reaction blocks provided by this 
invention have been demonstrated to be especially effective. Procedures 
for the release and isolation of nucleic acids can now be performed more 
quickly with this system than before. The system also makes it possible to 
leave the nucleic acids in the vessel during the steps described. This 
represents a considerable advance over the state of the art in terms of 
time savings and avoiding contamination. Suspensions were usually cooled 
previously by manually removing a sample processing vessel from the 
instrument and immersing it in a cooling bath. This procedure has been 
demonstrated to be insufficient for the future of routine diagnostic 
testing procedures. 
FIG. 3 shows a procedure for isolating nucleic acids provided by this 
invention. This figure is referred to in the procedure described in the 
example below. The sample vessel is located in a receptacle in unit 10. 
The sample vessel preferably has a stem (A20) that is molded to fit within 
the receptacle (e.g. it is conical in shape). The vessels shown in 
cross-section can be made simply from polypropylene using injection 
molding techniques. 
A main advantage of the invention is the fact that the system can be 
adapted to a large extent for use with different sizes of magnetic 
particles. It is relatively flexible and can be used with the most diverse 
procedures. 
The object of the invention is explained in greater detail using the 
example below: 
EXAMPLE 1 
The basic principles of the procedure provided by this invention are known 
by experts in nucleic acid diagnostics. Any technical details not listed 
below can be found in "Molecular Cloning" by J. Sambrook et al., CSH 1989. 
A particular construction of the procedure for processing sample solutions 
containing nucleic acids provided by this invention comprises the 
following working steps (see FIG. 3). In the initial step (I), a sample 
fluid containing cells is incubated in a sample vessel (A) with a material 
to which the cells bind and from which nucleic acids will be extracted. 
This material can have binding characteristics specific for cell surfaces, 
e.g. antibodies against surface antigens are immobilized on an absorber 
material (A16, not shown). Or, the material can have filter 
characteristics (A15, not shown) that retain the cells when the fluid 
flows through the material, e.g. as it is being removed from the sample 
vessel. Conditions for immobilizing cells on surfaces are known by the 
expert, e.g. from "Methods in Enzymology", Vol. 171, Biomembranes/Part R 
Transport Theory: Cell and Model Membranes, edited by Sidney Fleischer, 
Becca Fleischer, Department of Molecular Biology, Vanderbilt University, 
Nashville, Tenn., pages 444 ff or 581 ff. 
During incubation, the sample vessel is preferably closed with a cap (B) to 
ensure active and passive protection from contamination. 
In a subsequent step, the fluid is removed from the sample vessel while the 
cells containing the nucleic acids to be isolated remain in the sample 
vessel bound to the material. Since the cell-binding material is a 
particular material, retention is achieved by the fact that the material 
is magnetic (manufactured by Dynal, Oslo, Norway), and the magnet is moved 
towards the sample vessel from the outside. The fluid can be aspirated 
through the outlet (A11) by creating a slight vacuum. To accomplish this, 
a valve is built into the outlet that opens when a vacuum is created. 
One or more wash steps are provided in order to remove remaining sample 
components from the cells that may cause interference. In these steps, a 
wash fluid is added to the sample vessel in which any contaminants present 
dissolve but which does not have a negative effect on the cells binding to 
the surface of the cell-binding material. Wash solutions of this nature 
are known by the expert, e.g. from cell separation protocols or 
appropriate purification kit protocols for nucleic acids. They basically 
depend on the way the cells are bound to the material. 
Once the last wash solution is aspirated from the sample vessel (A), the 
purified, enriched cells are brought in contact with an appropriate lysis 
fluid to release the nucleic acids from the cells. The reagents of this 
lysis solution basically depend on the type of cells immobilized (Rolfs et 
al.: PCR, Clinical Diagnostics and Research, Springer Publishers, 1992, 
pg. 84 ff). If the cells are bacterial, the lysis solution preferably 
contains proteinase K to decompose the cell wall. If desired, the lysis 
can be encouraged with heating or cooling steps or mixing the reaction 
mixture by shaking the sample vessel. When lysis is complete, the nucleic 
acids to be isolated are freely available in the solution. 
The reaction vessel is also preferably closed with a cap during the lysis 
step, in order to prevent contamination from the environment. When lysis 
is complete, the cap is removed, preferably by using an appropriate 
mechanical device. A moulded article (C) is then inserted into the sample 
vessel that contains a mixture of cellular decomposition products and 
nucleic acids, the external contour of which (C12) is coordinated with the 
internal contour (A 17) of the sample vessel. This moulded article is 
hollow and closed with a filter (C 11) (porous matrix) on the end situated 
towards the sample vessel and the reaction mixture. The moulded article 
(C) is preferably inserted using a component (B 10) that is suitable for 
closing the sample vessel. In this case, the moulded article is held by 
the cap (II) and inserted into the sample vessel when it is closed. During 
this process, the reaction mixture also enters the hollow space (C 14) of 
the moulded article through the filter (C 11) (IV). The filter can prevent 
large particles from entering the hollow space and, due to its nucleic 
acid-binding characteristics, it binds nucleic acids even as the reaction 
mixture passes through. A filter material containing glass fibers is 
selected for use in this case. 
In a subsequent step, the remaining lysis reaction mixture is removed from 
the device formed by A and C by aspirating it through the outlet (All) in 
the sample vessel. This procedure also removes the solution that entered 
the hollow body (C14) of the moulded article, leaving as little filter 
residue in the filter as possible. The cap (B) is then removed, but the 
moulded article (C) remains in the sample vessel. 
At the same time or immediately thereafter, an elution vessel (D) is 
prepared to receive the moulded article (C) (either in the system provided 
by this invention or outside of it). If this vessel has a cap, it is 
removed (VI). Preferably, an elution solution is added to the elution 
vessel before the moulded article (C) is inserted into the elution vessel 
(D), e.g. using a pipette. The composition of the elution solution is 
based on the type of nucleic acid binding with the material in the filter 
(C). It contains reagents that cause the immobilized nucleic acids to 
elute, e.g. dissolve, from the material. The cap (B) that originally 
covered the elution vessel is placed on the sample vessel (A) with the 
moulded article (C) (VII). 
To remove the moulded article (C) from the sample vessel (A), the moulded 
article (C) is removed along with the cap (B) (VIII). The combination of 
cap and moulded article is then inserted in the elution vessel (IX). The 
moulded article (C) preferably contains a means (C13, not shown) for 
fixing the moulded article in the elution vessel (D) that ensures that the 
moulded article can only be removed from the vessel (D) if the moulded 
article (C) or vessel (D) is destroyed, or a force is used that is 
stronger than the force used to loosen the cap (B) from the moulded 
article (C). It is not intended for the moulded article to be removed from 
the elution vessel. 
While the moulded article (C) enters the elution vessel, the elution 
solution in the vessel enters the filter (C 11) and loosens the 
immobilized nucleic acids from the solid matrix. Depending on the quantity 
of elution solution in the vessel, the filter is either just saturated 
with the elution solution, or the elution solution--and the redissolved 
nucleic acids--enter the hollow space (C 14). To ensure that the nucleic 
acids are eluted as completely as possible, the inner contour of the 
elution vessel should fit as tightly against the external contour of the 
moulded article as possible. 
In a subsequent step, the cap (B) is removed from the combination of 
moulded article (C) and elution vessel (D) (X). It is used to pick up a 
stamp (E) (XI) and insert it into the hollow space of the moulded article 
(C) (XII). The cap grips the stamp (E) from inside. The stamp is pressed 
against the filter (C 11) with such force that fluid from the filter 
enters an internal space in the stamp through an opening on the pressing 
surface. This procedure is especially effective if the external contour of 
the pressing surface is coordinated with the inner contour of the moulded 
article (C), at least in the area intended for pressing. The stamp (E) can 
preferably be fixed in this position, e.g. by snapping it into place. 
Since the cap closes the device with this construction relatively tightly, 
the solution containing nucleic acids can be stored in this device. 
To remove a desired quantity of nucleic acid solution, the cap can be 
removed (XIII) and the desired quantity of solution can be removed through 
an opening in the internal space of the stamp, e.g. by pipette (XIV). The 
cap can then be placed back on the tube. 
The sequence of steps of the procedure described is provided below. 
______________________________________ 
Instrument Operator 
______________________________________ 
automated (program-controlled) 
manual 
incubate pipette 
aspirate place tubes, glass fleece insert and 
back-up container on the instrument 
separate (magnetic solid phase) 
mix/resuspend 
______________________________________ 
Manual working steps are shown in bold. Non-manual working steps or partial 
sequences are called up by pressing a key, for instance. 
In the table below, the sample vessel A is called a tube, the elution 
vessel D is referred to as the back-up vessel, the moulded article C is 
referred to as the glass fleece insert, and the stamp E is called the 
press stamp. 
______________________________________ 
Step 
# Action Time (s) 
______________________________________ 
1 Place tube #1-16 in the reaction module. 
2 Pipette receptor (50-100 .mu.l) and SA beads (50- 
100 .mu.l) in tubes #1-16 
3 Pipette sample (1000 .mu.l) in tubes #1-16 
4 Close tubes with cap (16 each) 
5 Mix. Frequency = 30 Hz (parallel) 
30 s 
6 Incubation. 9 = 4.degree. C. After incubation 9 = RT 
300-1200 s 
(parallel) 
6* Mix (if necessary) during incubation 
7 Magnet ACTIVATED (in-parallel) 
5 s 
8 Aspirate waste (sequentially, 5 s) 
80 s 
9 Magnet DEACTIVATED (in parallel) 
5 s 
10 Open the cap on the tubes (16 each) 
11 1st wash step: 
Pipette 500-1000 .mu.l wash solufion (low molecular 
weight salt) into tubes #1-16 
12 Resuspend. Frequency = 30 Hz (in parallel) 
5 s 
13 Magnet ACTIVATED (in parallel) 
5 s 
14 Aspirate waste (sequentially, 5 s) 
80 s 
15 Magnet DEACTIVATED (in parallel) 
5 s 
16 2nd wash step: 
Pipette 500-1000 .mu.l wash solution (low molecular 
weight salt) into tubes #1-16 
17 Resuspend. Frequency = 30 Hz (in parallel) 
5 s 
18 Magnet ACTIVATED (in parallel) 
5 s 
19 Aspirate waste (sequentially, 5 s) 
80 s 
20 Magnet DEACTIVATED (in parallel) 
5 s 
20.1 3rd wash step (optional): 
Pipette 500-1000 .mu.l wash solution (low molecular 
weight salt) into tubes #1-16 
20.2 Resuspend. Frequency = 30 Hz (in parallel) 
5 s 
20.3 Magnet ACTIVATED (in parallel) 
5 s 
20.4 Aspirate waste (sequentially, 5 s) 
80 s 
20.5 Magnet DEACTIVATED (in parallel) 
5 s 
21 Pipette lysis mix, reagent 1 + 2 (400 .mu.l) 
guanidinium hydrochloride or guanidinium 
rhodanide and proteinase K (25 .mu.l) in tubes #1-16 
22 Close the tubes with the caps (16 each) 
23 Resuspend. Frequency = 30 Hz (in parallel) 
5 s 
24 Incubation: 9 = 70.degree. C. 
600 s 
[Optional: Incubation: 9 = 95.degree. C. with potentially 
900 s 
infectious samples] 
25 Incubation: 9 = RT 300 s 
26 Open the caps on the tubes (16 each) 
27 Pipette 200 .mu.l ethanol (isopropanol) into tubes #1- 
16 
28 Close the caps bn the tubes (16 each) 
29 Mix. Frequency = 30 Hz (in parallel) 
30 Open the caps on the tubes (16 each) 
31 Place glass fleece insert in tube #1 into tubes #1-16 
32 Aspirate waste (sequentially, 5 s) 
80 s 
33 Pipette 500 .mu.l wash solution (chaotropic salt/ 
ethanol) into tubes #1-16 
34 Aspirate waste (sequentially, 5 s) 
80 s 
35 Pipette 500 .mu.l wash solution (chaotropic salt/ 
ethanol) into tubes #1-16 
36 Aspirate waste (sequentially, 5 s) 
80 s 
37 Place back-up vessel on the instrument (#1-16) 
38 Pipette elution volume into the back-up vessel 
(100-200 .mu.l) into tubes #1-16 
39 Transfer glass fleece insert from tube #1 to the 
back-up vessel (#1-16) 
40 Insert press stamp (#1-16) into the back-up vessel - 
Elution 
41 Close the back-up vessel (16 caps) 
42 Remove tubes #1-16 from the RM - waste 
______________________________________ 
If desired, the suction tubes and the cavities can be rinsed and, 
therefore, cleaned, with a cleaning fluid (before or after the procedure 
is performed and after the sample vessels have been removed). 
Another construction of a system for the release and isolation of nucleic 
acids is shown in FIGS. 4 through 8. 
FIG. 4: Perspective drawing of the system 
FIG. 5: Phantom drawing of the system 
FIG. 6: Receptacle for sample processing vessels 
FIG. 7: Cross-section through a receptacle for a sample vessel 
FIG. 8: Pushing device with magnets 
The system shown in FIG. 4 has 4 receptacles (100) for sample vessels. The 
receptacles (100) are fixed to a carrier (101). Moving the carrier (101) 
moves and shakes the 4 receptacles (110) in unison. The phantom drawing 
(FIG. 5) illustrates in greater detail how the movement of the carrier 
(101) takes place. The carrier has a circular recess (102) on each of its 
2 ends in which a rod (103) is situated. The rod (103) is situated on the 
axis of a motor (104) in an eccentric position. When the motor axis turns, 
therefore, the carrier is shifted along a plane. The phantom drawing (FIG. 
5) also illustrates clearly that the receptacle (100) has cooling fins 
(105). A stream of air is blown through the cooling ribs by means of 
ventilating fans (106) located on the base of the system to cool the 
system. 
A receptacle (100) is shown in greater detail in FIG. 6. The receptacle has 
6 separate cavities (107) to receive sample processing vessels. The 
cavities (107) are connected with the peltier element (109) by means of a 
framework (108) in a thermally conductive fashion. The peltier element, in 
turn, is connected with the cooling fins (105). The frame (108) contains 
temperature sensors and a resistance heating device to warm the cavities. 
The frame (108) is mounted on a peltier element (109) that exchanges heat 
with the cooling fins (105). The sample processing vessels are warmed by 
the heating elements located in the frame. The cavities (107) are cooled 
by the peltier element (109) in order to cool the sample processing 
vessels. The heat emitted by the peltier element on the opposite side is 
eliminated through the cooling fins (105). 
FIG. 7 shows a cross-section of one cavity (107). The cavity consists of a 
cylindrical space in which a sample vessel (110) is located. The sample 
vessel and cylindrical space are designed in such a way that the walls 
touch each other, to promote efficient heat transfer. Preferably, the 
sample vessel and cavity are both slightly conical, i.e. they taper 
towards the bottom, so that a snug fit is achieved. The conicity is 
preferably between 0.5 and 1.degree.. An insert is situated in the lower 
portion of the cavity that has a recess (112) on its underside into which 
a hose connection can be screwed. On the upper ends the insert (112) has 
an opening into which the tapered tip of a sample vessel (110) can be 
inserted. To achieve a tight connection between this opening and the 
tapered tip of the sample vessel, a sealing ring (113) in the shape of an 
O-ring that surrounds the tapered tip of the sample vessel is located in 
this position. To remove fluid from the sample vessel (110), a vacuum is 
created in the sample vessel (110) by means of a tube that is connected to 
the opening (112). 
When the system is operated as provided by this invention, a magnetic field 
can be applied to the sample vessels (110) while they are situated in the 
cavities (107), to hold magnetic particles. The magnet assembly shown in 
FIG. 8 is moved towards the cavities (107) for this purpose. 
FIG. 8 shows a moveable assembly of 4.times.6 magnets (114), corresponding 
to the 4 receptacles (110) with 6 cavities (107) each. The assembly shown 
in FIG. 8 has a rail (115) on which 4 units of 6 magnets each are mounted. 
The rail (115) is fixed to the carrier (116) which, in turn, holds a motor 
(117). A gearwheel (not shown) is mounted to the axis of the motor that 
grips the teeth (not shown) of a gear rack (118). The carrier (116) is 
arranged on cylinders (120) in such a fashion that it can be pushed along 
by linear ball bearings (119). In the position in which magnetic particles 
are deposited, the front surfaces of the magnets (114) are located 
directly against the flat sides (121) of the cavities (107). To move the 
magnets away from the cavities, the motor (117) is activated and the 
carrier, including the rail (115) is moved in linear fashion. 
______________________________________ 
Reference Drawing List 
______________________________________ 
A Sample Vessel 
10 Inlet 
11 Outlet 
17 Internal contour 
19 External contour 
20 Stem base 
22 Element to which additional functional elements can be 
attached 
B Cap 
10 Component for closing sample vessel A 
11 Component for gripping the moulded article C 
C Moulded Article 
11 Porous matrix 
12 External contour 
13 Means for flxing the moulded article in the elution vessel 
14 Hollow body 
15 Means for attaching a cap 
16 Internal contour 
17 Means for fixing a stamp E into position, all the way around 
18 Stem base, can be broken off 
19 Edge 
D Elution Vessel 
12 Snap-in notch 
E Stamp 
10 Pressing surface 
11 External contour 
12 Internal space 
13 Openings in the pressing surface 
14 Opening for removing contents 
15 Seal 
16 Snap-in ring 
17 Recess 
______________________________________ 
______________________________________ 
Instrument 
______________________________________ 
1 Frame 
10 Receptacle for sample vessels 
11 Vibration absorber 
12 Cavity 
13 Base 
14 Inlet to heat and cool A 
20 Thermostat unit to maintain sample vessels at a constant 
temperature 
21 Duct for cooling/heating 
30 Mechanical shaker for sample vessels 
40 Separator for separating magnetic particles using magnetic force 
41 Axis for turning magnet segments 
42 Magnet segments 
50 Vacuum pump/pump unit 
51 (Vacuum) tube 
100 Receptacle for sample vessels 
101 Carrier 
102 Circular recess 
103 Rod 
104 Motor 
105 Cooling fins 
106 Ventilator 
107 Cavity 
108 Frame 
109 Peltier element 
110 Sample vessel 
111 Insert 
112 Opening 
113 Sealing ring 
114 Magnet 
115 Rail 
116 Carrier 
117 Motor 
118 Gear rod 
119 Linear ball bearing 
120 Cylinder 
______________________________________