FlashGel™ System

Lonza Rockland, Inc.
www.lonza.com
scientific.support@lonza.com
Scientific Support: 800-521-0390
Customer Service: 800-638-8174
Rockland, ME 04841

Table of Contents

Safety Information

2

Cleaning and Disposal

4

Specifications

4

FlashGel™ System Quick Start Guide

6

FlashGel™ System for DNA Instructions

10

FlashGel™ System for RNA Instructions

19

FlashGel™ System for Recovery Instructions

25

FlashGel™ System Ordering Information

33

Warranty and Liability Information

37

License and Trademark Information

37

1

FlashGel™ System
Important Safety Information
Safety symbols
The following symbols alert the user to important operational,
maintenance, and/or warranty requirements, or possible hazards
exposure.

Symbol indicates a general caution or warning that the user may beaware of. Including but not limited to – potential chemical or hazardous
light exposure.

Symbol indicates a warning of potential exposure to hazardous voltage
where contact may result in death or serious injury.

General Product Warning Statement:
CAUTION: Hazardous Voltage
Contact may cause death or serious injury
Caution should be exercised in the operation of this system as it candevelop sufficient voltage and current to produce a lethal shock. To avoid
any risk of injury, the system should only be operated by properly trained
personnel and always in accordance with the instructions provided.
Prior to turning on the DC power source, ensure that the black lead isconnected to the negative terminal and the red lead is connected to thepositive terminal. Do not touch the FlashGel™ Dock or Cassette while thehigh voltage supply is turned on. Do not flood wells, add samples, or
extract bands while the high voltage leads are connected to the power
supply.
Failure to adhere to these instructions could result in personal and /orlaboratory hazards, as well as invalidate any warranty. Always turn off the
DC power source prior to removing cassettes from the dock. For maximum
safety, always operate this system in an isolated, low traffic area notaccessible to unauthorized personnel. Never operate damaged or broken
equipment.

2Precautions
The FlashGel™ Dock utilizes Dark Reader® Transilluminator technology(Clare Chemical Research, Inc.) to view fragments. It is safe to view
cassettes on the lighted dock without UV light protection. Turn on the light
only after the cassette is in place. Do not stare directly into the light.
CAUTION: Use the FlashGel™ Mask to block light from thesecond tier of wells when using double-tier or FlashGel™Recovery
Cassettes.
Wear gloves, lab coat, and safety glasses when handling FlashGel™
Cassettes. The gel and buffer in FlashGel™ Cassettes contain aproprietary nucleic acid gel stain that is a potential mutagen. Follow state
CAUTION: To avoid accidental exposure to high voltage, do notreverse electrodes in order to run samples backwards.
and local guidelines for handling and disposal of these materials.
Operating conditions for the FlashGel™ Dock
Maximum Limits
Electrophoresis unit: 0 - 300 VDC high voltage DC input
15 watts power
50 mA current
Dock light: 18 VDC low voltage DC input
Environmental Conditions
Operating Conditions: Temperature: 15C-35C Humidity: 15%-85% relative humidity, non-condensing For indoor use only
 Altitude up to 2000m
Storage and Shipping Conditions (FlashGel™ Dock) Temperature: 2C-60C Humidity: 15%-85% relative humidity, non-condensing

Cleaning, Maintenance and Disposal
3

Cleaning procedure
CAUTION: To avoid accidental exposure to high voltage, do notclean dock while connected to the high voltage power supply.
Clean the FlashGel™ Dock with a cloth moistened with water or mild
detergent. Do not immerse!

Maintenance
Visually inspect the dock prior to use for signs of wear, cracks or damage. Do notuse if damage is found.
There are no user serviceable parts contained in the FlashGel™ Dock.

Disposal
The stain in the FlashGel™ Cassettes is a potential mutagen. Follow state andlocal guidelines for disposal of these materials.

Specifications
Separation Range:
1.2% DNA Cassettes:
2.2% DNA Cassettes:
1.2% RNA Cassettes:
1.2% Recovery Cassettes:
2.2% Recovery Cassettes:

50 bp – 4 kb (up to 10 kb with longer run times)
10 bp – 1 kb
0.5 kb – 9 kb
50 bp – 4 kb
10 bp – 1 kb

Cassette Storage:
DNA Cassettes:
RNA Cassettes:
Recovery Cassettes:

18°C-26°C for 5 months from date of manufacture
Please inquire
18°C-26°C for 5 months from date of manufacture

Well Volume:
12 + 1 wells:
16 + 1 wells:
8 + 1 wells:

Do not exceed 5 l sample/well
Do not exceed 5 l sample/well
Do not exceed 12 l sample/wellGel Size:
70 mm (L) x 84 mm (W) x 2 mm (H)4

Cassette Size:
115 mm (L) x 107 mm (W) x 17 mm (H)Dock Size:
134 mm (L) x 120 mm (W) x 54 mm (H)Cassette Contents:
Agarose gel, stain and buffer

Equipment Ratings
Electrophoresis input (high voltage DC):
Voltage: 0 - 300 VDCPower: 15 W
Current: 50 mA
Dock light input (low voltage DC):
Voltage: 18 VDC
Current: 1.11 A
Dock light transformer input (line voltage AC):
Voltage: 100-240 VAC, 50-60 Hz
Current: 1.0 A
Electrical Connections:High voltage (electrophoresis): shielded, retractable banana plugs
Low voltage (light): 2.1 x 5.5 x 14 mm jackFlashGel™ Dock

High voltage cables forelectrophoresis

Low voltage power supply forviewing lightLight switch
Low voltage jack5

FlashGel™ System Quick Start Guide
Important Points
 Do not exceed 5 l sample volume per lane for 12 + 1 well and 16 + 1
well cassettes; or 12 l sample volume per lane for 8 + 1 wellcassettes.
 Optimal sample concentrations are approximately 1/5 the typical perband concentration of an ethidium bromide gel.
 For best results flood sample wells with water prior to loading, anduse FlashGel™ Loading Dye and FlashGel™ Markers, andFlashGel™ Recovery Buffer .
 Use the FlashGel™ Mask when running double-tier cassettes.
 Use the FlashGel™ Visualization Glasses when recovering samples.
Instructions
1. Refer to Table 1 (pp 8-9) for recommended sample preparation and run
conditions.
2. Remove white well seals from cassette. Do not remove the clear sidevent seals.
3. Flood sample wells with distilled or deionized water. Tilt cassette tomove excess fluid to the edge and blot off with a lint free wipe. Do notblot wells directly.
4. Insert cassette into dock. Insert FlashGel™ Mask under the central tierof sample wells if using double-tier or recovery cassettes.
5. Load samples. Samples to be recovered should be loaded in upper tiersample wells.

66. Plug in high voltage cables, turn on power supply and set torecommended voltage.
7. Plug in low voltage power supply and turn on light.
8. If using standard DNA and RNA cassettes run for recommended time oruntil separation of desired fragments is complete. If using recovery
cassettes run and observe migration of sample; just prior to desired
sample reaching recovery wells (2nd-tier) stop the run and disconnect
the high voltage cables (complete steps 9 – 13).
9.

Blot excess buffer from the recovery well(s) and add 20 l of FlashGel™
Recovery Buffer.

10. Remove FlashGel™ Mask, reconnect voltage cables, and restart power.
Use FlashGel™ Visualization Glasses to observe band migration.
11. When the band of interest has migrated to the center of the recovery
well, turn off power supply and disconnect voltage cables. Use a pipette
to carefully remove the recovery buffer (containing the DNA) from therecovery well.
12. If necessary, the process (addition of recovery buffer, electrophoresis,
and recovery) may be repeated to increase recovery of higher DNA
loads.
13. Photograph using The FlashGel™ Camera, or other standard
camera and transilluminator.

7

Table 1. Recommended Sample Preparation and Run Conditions
DNA Cassettes

RNA Cassettes

Recovery Cassettes

Separation range

1.2%: 50 bp – 10 kb
2.2%: 10 bp – 1 kb
Separation of fragments> 4 kb will be improvedby running longer atlower voltage

1.2%: 0.5 kb – 9 kb

1.2%: 50 bp – 10 kb
2.2%: 10 bp – 1 kb
Separation of fragments> 4 kb will be improvedby running longer atlower voltage

Samplepreparation

For best results, dilute
DNA samples in 1X
FlashGel™ Loading Dye

Denatured RNA
samples: Prepare
samples in 50%
formaldehyde
sample buffer and
RNase-free water;
denature for 5
minutes at 65°C.

For best results, dilute
DNA samples in 1X
FlashGel™ Loading Dye

Sampleconcentration anddetection limits

Optimal DNA load levels
are 5-20 ng/band in a 5
l load; for best results,
do not exceed 20
ng/bandVoltage & run timeSingle-tier: 275 V for 2-7
minutes

Native RNA
samples: Use
FlashGel™ Loading
Dye
Optimal RNA load
levels will varydepending upon
RNA sample; forbest results, do not
exceed 200
ng/band in a 5lloadSingle-tier: 225 V
for 4-8 minutes

Double-tier: 275 V for 2-5
minutes

Double-tier: 225 V
for 3-5 minutes

N/A

N/A

Recoveredconcentration andvolume8

Optimal DNA load levels
are 50-500 ng/band inload volumes up to 12 l

275 V for time needed toelectrophorese bands torecovery wells – varies
by fragment size from 3+
minutes
Maximum run time 12-14
minutes
Sample recoveries are
typically 80-90%,
depending upon
fragment
Recovery volumes are
typically 15-50lRecommended
Markers

DNA Cassettes

RNA Cassettes

1.2% cassettes:
FlashGel™ DNA Marker
100 bp–4 kb

FlashGelTM RNA
Marker 0.5 kb–9 kb

Recovery Cassettes
FlashGel™ DNA
Marker 100 bp–3 kb
FlashGel™
QuantLadder
100 bp-3 kb

2.2% cassettes:
FlashGel™ DNA Marker
50 bp – 1.5 kb

2.2% cassettes:
FlashGel™ DNA
Marker 50 bp – 1.5 kb

Double-tier cassettes:
FlashGel™ DNA Marker
100 bp – 3 kb

9

FlashGel™ System for DNA Instructions
Introduction
The FlashGel™ System is recommended for fast separation and analysis
of DNA.
FlashGel™ DNA Cassettes:
 Sizing and/or quantitation of PCR or restriction fragments 10 bp – 10 kb
 Confirmation of PCR amplification
Separation of DNA fragments can be monitored in real time at the labbench without the use of UV illumination.
Fragments separated with the FlashGel™ System may be photographedwith The FlashGel™ Camera, or other standard documentation systems.
FlashGel™ DNA Cassettes are not recommended for recovery. Use
FlashGel™ Recovery Cassettes (page 25) for DNA to be recovered.
Important Information
Run conditions and resolution
The FlashGel™ System is designed for fast, high voltage separation offragments 10 bp to 10 kb. Cassettes may be run at lower voltages forlonger times to improve separation of fragments > 4 kb (page 18). Monitor
the run and optimize conditions. Fragments will run faster on a warm
dock. Reduce run time, or lower voltage as necessary. Refer to Table 1(pp 8-9) for recommended run conditions.
Visualization of bands on the FlashGel™ Dock
DNA and marker bands will be visible on the lighted dock under typical lab
lighting. The degree of visibility may vary based upon the overall lightintensity in the lab. Bands are best viewed looking straight down at thedock in a space that minimizes light intensity and reflection.
Alternatively, band separation may be visualized on a computer screen withthe aid of the FlashGel™ Camera.
To ensure adequate visibility of a marker for monitoring the run on thedock, use FlashGel™ DNA Markers or FlashGel™ QuantLadders. Refer to10

Table 2 (pp 14) and Figure 2 (page 16) for details on DNA concentrations
visible under different conditions.
DNA bands are visible on the dock throughout the run.
Loading dye and markers
The FlashGel™ System is compatible with standard gel loading dyes andmarkers. Bromophenol blue dye does not migrate in FlashGel™ Cassettes;
however samples containing bromophenol blue may be used, as sample
migration is not affected. Use the FlashGel™ Loading Dye and FlashGel™
Markers or FlashGel™ QuantLadder for best results.
Preparing the gel for running
For best results, use a transfer pipette or squirt bottle to flood sample wellswith distilled or deionized water before loading samples or markers. This
will ensure adequate moisture in the wells. Flood the wells, then tilt thecassette and use a lint free wipe or small piece of blotting paper to remove
excess liquid from the cassette. Do not blot the wells directly.
DNA sensitivity levels
The FlashGel™ System uses a proprietary stain that is 5–20 times moresensitive than ethidium bromide stain. In general, use per band DNA
concentrations 1/5 of those normally used for ethidium bromide detection.
Optimal DNA load levels on FlashGel™ Cassettes are 5-20 ng per band.
DNA levels below 5 ng per band may not be visible on the dock, but levels
as low as 0.10 ng per band can be detected on gel images/photos. DNA
levels up to 80 ng per band can be used; however, levels exceeding 20 ng
per band may result in band distortion. DNA levels may be adjusted toprovide best performance for the image analysis system used.
Because of the sensitivity of the FlashGel™ System, most DNA samples
should be diluted, and load volume per well must not exceed 5 l.
For best results, dilute DNA samples into 1X FlashGel™ Loading Dye suchthat a 5 l sample contains 5-10 ng DNA per band.

11Gel Running Instructions
CAUTION: Wear gloves, lab coat and safety glasses whenhandling FlashGel™ Cassettes.
1. Prepare samples. Refer to Table 1 (pp 8-9) for recommended sample
preparation.
2. Tear open the pouch and remove the cassette.
NOTE: If cassette is wet, dry with a clean wipe.
NOTE: Air pockets may be present between the gel and cassette.
These will not affect band migration.
3. Place the cassette on a flat surface and pull the tab to remove the white
well seal. Do not remove the clear seals covering the side vent holes.
4. Flood all sample wells with distilled or deionized water. Tilt the cassetteand use a lint free wipe or small piece of blotting paper to remove
excess fluid. Do not blot the wells directly.
CAUTION: To avoid accidental exposure to high voltage, do not
flood wells while cassette is connected to the high voltage power
supply.
NOTE: For best results, flood wells with AccuGENE™ Molecular
Biology Water (Cat. #51200).
5. Place the cassette on the dock and slide it into place; the cassetteshould snap securely into the dock.
6. Load samples and markers. Refer to Table 1 (pp 8-9) for sample
preparation and recommended markers.

12

CAUTION: To avoid accidental exposure to high voltage, do notload samples while cassette is connected to the high voltage power
supply.
7. Connect the low voltage power supply to the dock by inserting the lead
into the receptacle at the back of the dock and plugging in the power
supply. This is the power supply to the light source.
8. Turn on the dock light by pressing the orange button on the top of theunit.
NOTE: The light will automatically shut off after 10 minutes. Re-start bypressing the orange button.
9. Connect the high voltage leads to power supply and set power atrecommended voltage (Table 1, pp 8-9).
NOTE: Typical starting currents should range from 20-25 mA for DNA
gels. Run until desired separation is obtained for the fragments ofinterest. Under the conditions outlined, DNA fragments of 50 bp-100 bp
will migrate out of the gel in 7-8 minutes. Refer to Figure 1 (page 15) for
separation results at various run times.
NOTE: Running multiple cassettes in quick succession may requireslight adjustment of running conditions as bands will run faster as thedock gets warmer. Monitor separation and reduce voltage and run time ifnecessary.
NOTE: Some expression of buffer from the wells is normal.
CAUTION: Handle cassettes only after voltage is turned off andleads are disconnected.

10. Run DNA cassettes until desired separation is reached for the fragmentof interest.

1311. Turn off voltage and disconnect leads from the power supply prior to
removing the cassette.
12. Record the gel data using The FlashGel™ Camera, by Polaroid®
photography, or by other image capture systems. See Figure 2 (page16) for details on typical cameras.

Reference Information
Table 2. DNA Amount Visible on FlashGel™ DNA Cassettes Under
Various Conditions

Viewed in ambient light

1500 bp Fragment
1.6 – 3.2 ng

400 bp Fragment
1.6 – 3.2 ng

Viewed in Dark Room

0.39 – 0.78 ng

0.39 – 0.78 ng

Viewed on FlashGel™
Camera0.10 – 0.20 ng

0.10 – 0.20 ng

FlashGel™ Camera
Photo

0.10 – 0.20 ng

0.10 – 0.20 ng

Viewed on Dark Reader®
Transilluminator

0.05 – 0.01 ng

0.10 – 0.20 ng

Dark Reader® Photo

0.10 – 0.20 ng

0.10 – 0.20 ng

Viewed on UV

0.39 – 0.78 ng

0.39 – 0.78 ng

UV Photo

0.39 – 0.78 ng

0.39 – 0.78 ng

14

Fig 1. Examples of Separation at Various Run Times at 275 V on aFlashGel™ DNA Cassette (1.2%, 12+1 well single-tier)2 minutes

4 minutes

8 minutes

6 minutes

10 minutes

Sample lanes left to right:
Lane 1. FlashGel™ DNA Marker 100/200/300/500/800/1250/2000/4000 bp(5 l load 1:5 dilution)
Lane 2. FlashGel™ QuantLadder 100/250/400/800/1500 bp(5 l load 1:5 dilution)
Lane 3. Lonza Marker 50 bp – 2500 bp (3 l load 1:5 dilution)
Lane 4. Lonza Ladder 100 bp (3 l load 1:15 dilution)
15

Fig 2. Detection of DNA Fragments on FlashGel™ DNA Cassettes






Illuminate cassettes using UV or blue-light transilluminators, such as
the Dark Reader® Transilluminator.
Photograph using FlashGel™ Camera, or any system used for standard
ethidium bromide stained gels.
For Polaroid® Type 57 film with a UV transilluminator, start with 1-2
second exposures.
For CCD systems, use the system’s existing ethidium bromide filter.
FlashGel™ DNA Marker (100 bp - 4 kb) is loaded in far right lane.

Images below show a dilution series of 400 bp and 1500 bp fragments
Dark Reader® Transilluminator with orange cover, CCD Camera with EtBr Filter,3 second exposure
75

50

25

12.5 6.25 3.13 1.56 0.78 0.39 0.20 0.10 0.05 ng/lane

UV transilluminator, Polaroid® Camera with EtBr Filter, 3 second exposure
75

50

25 12.5 6.25 3.13 1.56 0.78 0.39 0.20 0.10 0.05 ng/lane

16

Fig. 3. Separation of Markers and DNA Fragments on a FlashGel™
DNA Cassette (2.2%, 16 + 1 well double-tier)Sample lanes:
Lanes 1, 7, 13: FlashGel™ 50 bp – 1.5 kb Marker(recommended for 2.2% cassettes)
Lanes 2, 8, 14: FlashGel™ 100 bp – 3 kb Marker(recommended for double-tier cassettes)
Lanes 6 and 12: FlashGel™ QuantLadder
Lanes 3, 4, 5/9, 10, 11/15, 16, 17: 8 ng loads of 150 bp, 300 bp and 800 bp
DNA fragments

17

Fig. 4. Improving Large DNA Fragment Separation on the FlashGel™ System
2 – 4 kb

3 - 10 kb

Fragment sizes:
2000/2500/3000/4000 bp

Fragment sizes:
3000/4000/5000/7000/10000 bp

18

FlashGel™ System for RNA Instructions
Introduction
The FlashGel™ System is recommended for fast separation and analysisof RNA, including: Verification and analysis of total RNA 0.5 kb – 9 kb
 Checking for RNA degradation
 Quick checks of native RNA
Fragments separated with the FlashGel™ System may be photographedusing the FlashGel™ Camera or other standard documentation systems.
FlashGel™ RNA Cassettes are not recommended for recovery.
Important Information
Run conditions and resolution
The FlashGel™ System is designed for fast, high voltage separations.
Cassettes may be run at lower voltages for longer times to improveseparation of higher molecular weight fragments. Monitor the run andoptimize conditions. Fragments will run faster on a warm dock. Reducerun time, or lower voltage as necessary. Refer to Table 1 (pp 8-9) forrecommended run conditions.
Visualization of bands on the FlashGel™ Dock
RNA and marker bands will be visible on the lighted dock under typical lab
lighting. The degree of visibility may vary based upon the overall lightintensity in the lab. Bands are best viewed looking straight down at thedock in a space that minimizes light intensity and reflection.
RNA bands will be visible on the dock for the first 3-4 minutes of the run,after which they fade and then reappear after a > 10 minute post-run holdperiod. The FlashGel™ DNA Marker may be used to monitor the RNA run.
Loading dye and markers
The FlashGel™ System for RNA is compatible with standard formaldehyde
loading dye and RNA markers. Bromophenol blue dye does not migrate inFlashGel™ Cassettes; however samples containing bromophenol blue may

19be used, as sample migration is not affected. Use the Lonza
Formaldehyde Sample Buffer or FlashGel™ Loading Dye (for native RNA),and FlashGel™ RNA Marker for best results.
Preparing the gel for running
For best results, use a transfer pipette or squirt bottle to flood sample wellswith RNase-free water before loading samples or markers. This will ensure
adequate moisture in the wells. Flood the wells, then tilt the cassette anduse a lint free wipe or small piece of blotting paper to remove excess liquidfrom the cassette. Do not blot the wells directly.
RNA sensitivity levels
The FlashGel™ System uses a proprietary stain that is 5–20 times moresensitive then ethidium bromide stain and will detect RNA quantities < 10ng per band.
For denatured RNA, dilute samples with formaldehyde loading buffer suchthat a 5 l load contains < 200 ng RNA per band.
For native RNA, dilute samples into FlashGel™ Loading Dye such that a
5 l load contains < 200 ng RNA per band.
Gel Running Instructions
CAUTION: Wear gloves, lab coat and safety glasses whenhandling FlashGel™ Cassettes.

1. Prepare samples. Refer to Table 1 (pp 8-9) for recommended sample
preparation.
2. Tear open the pouch and remove the cassette.
NOTE: If cassette is wet, dry with a clean wipe.
NOTE: Air pockets may be present between the gel and cassette.
These will not affect band migration.

20

3. Place the cassette on a flat surface and pull the tab to remove the white
well seal. Do not remove the clear seals covering the side vent holes.
4. Flood all sample wells with RNase-free water. Tilt the cassette and use
a lint free wipe or small piece of blotting paper to remove excess fluid.
Do not blot the wells directly.
NOTE: For best results, flood wells with AccuGENE™ Molecular
Biology Water (Cat. #51200).
CAUTION: To avoid accidental exposure to high voltage, do not
flood wells while cassette is connected to the high voltage power
supply.

5. Place the cassette on the dock and slide it into place; the cassetteshould snap securely into the dock.
CAUTION: To avoid accidental exposure to high voltage, do notload samples while cassette is connected to the high voltage power
supply.

6. Load samples and markers. Refer to Table 1 (pp 8-9) for sample
preparation and recommended markers.
CAUTION: Use the FlashGel™ Mask to block light from thesecond tier of wells when using double-tier cassettes.
7. Connect the low voltage power supply to the dock by inserting the lead
into the receptacle at the back of the dock and plugging in the power
supply. This is the power supply to the light source.
8. Turn on the dock light by pressing the orange button on the top of theunit.
NOTE: The light will automatically shut off after 10 minutes. Re-start bypressing the orange button.
21

9. Connect the high voltage leads to power supply and set power atrecommended voltage (Table 1, pp 8-9).
NOTE: Typical starting currents should be 25 - 30 mA. Run until desired
separation is obtained for the fragments of interest.
NOTE: Running multiple cassettes in quick succession may requireslight adjustment of running conditions as bands will run faster as thedock gets warmer. Monitor separation and reduce voltage and run time ifnecessary.
NOTE: Some expression of buffer from the wells during run is normal.
10. Run RNA cassettes for 8 minutes, then turn off voltage and disconnect
leads from the power supply prior to removing the cassette.
CAUTION: Handle cassettes only after voltage is turned off andleads are disconnected.
11. Remove the cassette and let it stand at room temperature for >10
minutes, or until the fragments are visible at the desired intensity.
Maximum intensity is reached in approximately 45 minutes.
Record the gel data using The FlashGel™ Camera, or by Polaroid®
photography or other image capture systems. See Figures 5 and 6 (pp23-24) for RNA images.

22Reference Information
Fig. 5. Detection of RNA Fragments on a FlashGel™ RNA Cassette
1

2

3

4

5

6

7

8

9

10

11

12 13

Cassette run for 8 minutes at 225 V; photographed 20 minutes post-run.
Sample Lanes:
Lanes 1, 6, 11: DNA Marker (for visualization during run)Lanes 2, 7, 12: 100 ng RNA Ladder
Lanes 3, 8, 13: 100 ng E. coli Total RNA (Ambion)
Lanes 4, 9: ~100 ng S. cervevisiae Total RNA purified using RiboPure™
Yeast Kit (Ambion)
Lanes 5, 10: 100 ng Mouse Thymus Total RNA (Ambion)

23

Fig. 6. Quick Check of Native RNA Fragments on a FlashGel™ RNA
Cassette (1.2%, 12+1 single-tier)
1

2

3

4

Cassette run for 4 minutes at 225 V; followed by immediate imaging.
Sample lanes:
Lane 1: Lonza Marker, 50 ng
Lane 2: E. Coli Total RNA, 50 ng
Lane 3: Lonza Marker, 250 ng
Lane 4: E. Coli Total RNA, 250 ng

24FlashGelTM System for Recovery Instructions
Introduction
The FlashGel™ System for Recovery is recommended for fast separation
and recovery of sheared DNA and fragments from 10 bp – 4 kb.
Separation of DNA fragments can be monitored in real time and fragments
can be recovered at the lab bench without the use of UV illumination, orthe need for band excision.
Fragments separated with the FlashGel™ Recovery System arecompatible with standard molecular biology applications (PCR
amplification, ligation and cloning, etc.)
Important Information
Run conditions and resolution
The FlashGel™ System for Recovery is designed for fast, high voltage
separations and recovery. Cassettes may be run at lower voltages forlonger times to improve separation of larger fragments. Monitor the run
and optimize conditions for the fragment of interest. Fragments will run
faster on a warm dock. Reduce run time, or lower voltage as necessary.
Do not exceed 14 minutes total run time. Refer to Table 1 (pp 8-9) forrecommended run conditions.
Visualization of bands on the FlashGel™ Dock
DNA and marker bands will be visible on the lighted dock under typical lab
lighting. The degree of visibility may vary based upon the overall lightintensity in the lab. Bands are best viewed looking straight down at thedock in a space that minimizes light intensity and reflection.
To ensure adequate visibility of a marker for monitoring the run, use
FlashGel™ DNA Markers or FlashGel™ QuantLadders. Refer to Table 2(page 14) for details on DNA concentrations visible under different
conditions.
Loading dye and markers
The FlashGel™ System is compatible with standard gel loading dyes andmarkers. Bromophenol blue dye does not migrate in FlashGel™
Cassettes; however samples containing bromophenol blue may be used,25

as sample migration is not affected. Use the FlashGel™ Loading Dye andFlashGel™ Markers or FlashGel™ QuantLadder for best results.
Preparing the gel for running
For best results, use a transfer pipette or squirt bottle to flood sample wellswith distilled or deionized water before loading samples or markers. This
will ensure adequate moisture in the wells. Flood the wells, then tilt thecassette and use a lint free wipe or small piece of blotting paper to remove
excess liquid from the cassette. Do not blot the wells directly.
For best results, dilute DNA sample into 1X FlashGel™ Loading Dye so thesample (12 l maximum load) contains the suggested amount of DNA perband.
DNA sensitivity levels
The FlashGel™ System uses a proprietary stain that is 5–20 times moresensitive than ethidium bromide stain. In general, use per band DNA
concentrations 1/5 of those normally used for ethidium bromide detection.
Optimal DNA load levels on FlashGel™ Recovery Cassettes are 50-500 ng
per band. DNA levels may be adjusted to provide the best performance forthe fragments of interest. High DNA load levels may not provide adequate
resolution if the fragment of interest is relatively close in size to othercontaminating fragments.
DNA Recovery
For optimal recovery efficiency (particularly of larger DNA fragments) add
FlashGel™ Recovery Buffer to the Recovery wells prior to extracting DNA
fragments.
To identify optimal conditions for recovered fragments use the FlashGel™
Control Fragment provided in the FlashGel™ Recovery Starter Pack.
Resolution of Recovered DNA
For best resolution of recovered DNA, dilute the sample to be run on asubsequent gel at least 1:1 with water (e.g. 2 l recovered DNA, 2 l water,
1 l 5X FlashGel™ Loading Dye).

26

Gel Running Instructions
CAUTION: Wear gloves, lab coat and safety glasses whenhandling FlashGel™ Cassettes.
1. Prepare samples. Refer to Table 1 (pp 8-9) for recommended sample
preparation.
2. Tear open the pouch and remove the cassette.
NOTE: If cassette is wet, dry with a clean wipe.
NOTE: Air pockets may be present between the gel and cassette.
These will not affect band migration.
3. Place the cassette on a flat surface and pull the tab to remove the white
well seal. Do not remove the clear seals covering the side vent holes.
4. Flood both tiers of wells with distilled or deionized water. Tilt thecassette and use a lint free wipe or small piece of blotting paper toremove excess fluid. Do not blot the wells directly.
NOTE: For best results, flood wells with AccuGENE™ Molecular
Biology Water (Cat. #51200).
CAUTION: To avoid accidental exposure to high voltage, do not
flood wells while cassette is connected to the high voltage power
supply.

5. Place the cassette on the dock and slide it into place; the cassetteshould snap securely into the dock. Slide the FlashGel™ Mask in place
under the second tier of wells to minimize light passing through the wells
and increase the ease of viewing as samples separate.
CAUTION: To avoid accidental exposure to high voltage, do notload samples while cassette is connected to the high voltage power
supply.
27

CAUTION: Use the FlashGel™ Mask to block light from the second
tier of wells when using Recovery Cassettes.
6. Load samples and markers. Refer to Table 1 (pp 8-9) for sample
preparation and recommended markers.
NOTE: Use FlashGel™ Control Fragment to monitor recovery.
7. Connect the low voltage power supply to the dock by inserting the lead
into the receptacle at the back of the dock and plugging in the power
supply. This is the power supply to the light source.
8. Turn on the dock light by pressing the orange button on the top of theunit.
NOTE: The light will automatically shut off after 10 minutes. Re-start bypressing the orange button.
CAUTION: Handle or touch cassettes only after voltage is turned
off and leads are disconnected.

9. Connect the high voltage leads to power supply and set power atrecommended voltage (Table 1, pp 8-9).
NOTE: Typical starting currents should range from 20-25 mA.
NOTE: Some expression of buffer from the wells during run is normal.
Wear gloves, lab coat and safety glasses when handling.
10. Allow run to proceed, monitoring migration of sample(s) to be recovered.
Just prior to desired sample(s) reaching recovery wells (2nd tier),
turn off power and disconnect the high voltage cables. Do notremove the cassette from the dock.
CAUTION: Handle or touch cassettes only after voltage is turned
off and leads are disconnected.

28

11. Blot excess buffer from the recovery well(s) and add 20 l/well ofFlashGel™ Recovery Buffer.
12. Remove FlashGel™ Mask, reconnect voltage cables, and restart power.
Use FlashGel™ Visualization Glasses to observe DNA band migration.
13. Run until the band to be recovered has entered the recovery well, andthe leading edge of the band has moved to the front edge of therecovery well.
NOTE: When recovering more than one fragment from gel, recoversmallest fragment first.
14. Turn off power, disconnect leads, and then use a pipette to remove
the recovery buffer containing the DNA.
CAUTION: Wear gloves, lab coat and safety glasses whenhandling FlashGel™ Cassettes.
NOTE: The amount recovered will not be the full 20 µl loaded into thewell.
15. For recovery of large quantities of DNA (>350 ng), it may be necessary
to repeat the electrophoresis-recovery cycle to retrieve the maximum
DNA material.
If necessary, add an additional 20 l of FlashGel™ Recovery Buffer andstart the run again until the sample has once again moved to the frontedge of the recovery well, then recover. Repeat as necessary, being
careful to always turn off the power and disconnect the leads prior
to recovering samples.
NOTE: The buffer capacity of the FlashGel™ Recovery Cassette will
support ~12-14 minutes of total run time at 275 V. Use care in recovery
of multiple fragments or exceptionally large fragments to avoid excess
run time.

29

NOTE: It may be necessary to repeat steps 10 – 15 when recovering
sheared DNA if your size selection is wider than the recovery well.
Utilize the different cassette concentrations available to optimize yoursize selection needs.
16. Sample recoveries may be quickly assessed using the standard
FlashGel™ Markers for DNA and/or the FlashGel™ QuantLadder.
Prepare sample of recovered DNA by combining equal volumes ofrecovered DNA and water then adding appropriate amount of 5X
FlashGel™ Loading Dye. The volume of DNA sample for recovery
check depends upon the amount of input DNA loaded on the recovery
gel. Estimate the amount of DNA in the recovered sample bycomparison to bands in the FlashGel™ QuantLadder (Table 3, page 31;
Fig. 8, page 32).
NOTE: Depending upon the amount of run time used for the recovery
step, the same FlashGel™ Cassette may be used to check therecovered sample. The cassette will be functional up to ~ 12-14 minutesof total run time at 275 V.

30

Reference Information
Table 3. FlashGel™ QuantLadder DNA Levels
Fragment
1500 bp

2.5 µl Load15 ng

5.0 µl Load
30 ng

800 bp10.5 ng

21 ng

400 bp

7.5 ng

15 ng

250 bp3.75 ng

7.5 ng100 bp

1.5 ng3 ng

Fig. 7. Images of FlashGel™ Recovery Cassettes Before and After
Recovery
BeforeAfter1000 bp DNA Fragment(300 ng load)

After

Samples from two lanes removed from a 2.2% FlashGel™
Recovery Cassette. Several seconds of 275V current applied
after samples recovered show selection removed. Samples ofhigher molecular weight are now ready to be recovered.

31

Fig. 8. Recovery of Wide Size Range of DNA Fragments on FlashGel™
DNA Cassette

Samples (100 ng) of DNA fragments were separated andrecovered using the FlashGel™ Recovery System. Image
above shows the analysis of 3 l aliquots of recovered
DNA using a 1.2% DNA FlashGel™ Cassette.

Recovered samples of various size
selections from fragmented genomic
lambda DNA. Lanes contain DNA
extractions from several experiments takenfrom 1.2% FlashGel™ Recovery Cassettes.

Sample Lanes:

Sample Lanes:

Lane 1: FlashGel™ DNA Marker 100-4000 bp

Lane 1: 280 – 550 bp*

Lane 2: FlashGel™ QuantLadder (2.5 µL)

Lane 2: 650 – 1,190 bp*

Lane 3: 50 bp DNA fragment

Lane 3: 425 – 800 bp*

Lane 4: 200 bp DNA fragment

Lane 4: 425 – 800 bp*(smaller load volume of lane 3)

Lane 5: 500 bp DNA fragment
Lane 5: 800 – 3,000 bp*
Lane 6: 1000 bp DNA fragment*Size ranges were determined bygel electrophoresis

Lane 7: 2000 bp DNA fragment
Lane 8: 4000 bp DNA fragment

32

Ordering Information
FlashGel™ System
Cat No.
57025

Description
FlashGel™ Dock

Size/Format
One size

57040

FlashGel™ Camera

One size

57068

FlashGel™ Power Supply

One size

57062

FlashGel™ Device Pack

57069

FlashGel™ Power Supply Pack

57065

FlashGel™ Camera Pack

Includes FlashGel™ Dock,
FlashGel™ Power Supply, andFlashGel™ Camera
Includes FlashGel™ Dock andFlashGel™ Power Supply
Includes FlashGel™ Dock andFlashGel™ Camera

FlashGel™ System for DNA
Cat No.

Description

Size/Format

57023

FlashGel™ DNA Cassettes

1.2% agarose,
12+1 well single-tier format, 9pk

57029

FlashGel™ DNA Cassettes

1.2% agarose,
16+1 well double-tier format, 9pk

57031

FlashGel™ DNA Cassettes

2.2% agarose,
12+1 well single-tier format, 9pk

57032

FlashGel™ DNA Cassettes

2.2% agarose,
16+1 well double-tier format, 9pk

50462

FlashGel™ Loading Dye

5 x 1 ml vials
5X concentration

50473

FlashGel™ DNA Marker
100 bp – 4 kb

500 l Ready-to-load
Recommended for 1.2% cassettes
Band Sizes:
100/200/300/500/800/1250/2000/4000
bp

33

FlashGel™ System for DNA (cont.)
Cat No.
57033

Description
FlashGel™ DNA Marker
50 bp – 1.5 kb

Size/Format
500 l Ready-to-load
Recommended for 2.2% cassettes
Band Sizes:
50/100/150/200/300/500/800/1500 bp

57034

FlashGel™ DNA Marker
100 bp – 3 kb

50475

FlashGel™ QuantLadder

57026

FlashGel™ DNA Kit

500 l Ready-to-load
Recommended for double-tier cassettes
Band Sizes: 100/300/500/800/1500/3000 bp
250 l Ready-to-load
Band Sizes: 100/250/400/800/1500
Includes FlashGel™ DNA Cassettes
1.2% 12+1 well single tier 9pk,
FlashGel™ Loading Dye, andFlashGel™ Marker 100 bp – 4 kb

FlashGel™ System for RNA
Cat No.
Description
57027
FlashGel™ RNA Cassettes

Size/Format
1.2% agarose,
12+1 well single-tier format, 9pk

57028

FlashGel™ RNA Cassettes

1.2% agarose,
16+1 well double-tier format, 9pk

50571

Formaldehyde Sample Buffer

RNA denaturing sample buffer,contains bromophenol blue andxylene cyanol, 5 x 1 ml

50462

FlashGel™ Loading Dye

RNA native sample buffer
5 x 1 ml vials
5X concentration

50577

FlashGel™ RNA Marker

0.5 bp – 9 kb
50 µg (1 µg/ml)

51200

AccuGENE™ Molecular Biology For flooding sample wells andWater (DNase/RNase free)diluting RNA, 1L

34

FlashGel™ System for RNA (cont.)
Cat No.
Description
57024

FlashGel™ RNA Kit

Size/Format
Includes FlashGel™ RNA Cassettes
1.2% 12+1 well single tier 9pk, RNA
Marker, Formaldehyde Sample Buffer,and Molecular Biology Water

FlashGel™ System for Recovery
Cat No.
57051

Description
Size/Format
FlashGel™ Recovery Cassettes 1.2% agarose
8 + 1 double-tier format, 9pk

57022

FlashGel™ Recovery Cassettes 2.2% agarose
8 + 1 double-tier format, 9pk

57060

FlashGel™ Recovery Buffer

2 x 500 l vials

57050

FlashGel™ Recovery Kit

Includes FlashGel™ Recovery
Cassettes 1.2% 8+1 well doubletier 9pk, FlashGel™ Recovery
Buffer, FlashGel™ Loading
Dye, FlashGel™ QuantLadder,and Visualization Glasses

35Notes36

Warranty and Liability
This product was produced utilizing the highest practical standards of materials, workmanship,and design. Lonza Rockland, Inc. warrants that the product has been tested and will meet or
exceed published specifications. This warranty is valid only if the product has been operated
and maintained according to the instructions provided.
Lonza Rockland, Inc. warrants this product to be free from defects in materials and workmanshipunder normal service for one year from date of shipment. If the product proves defective during
this period, Lonza Rockland, Inc., will repair or replace it at our option, free of charge, if
returned to us postage prepaid. This warranty does not cover: damage in transit, damage caused
by carelessness, misuse or neglect, normal wear through frequent use, damage caused by solvent
corrosion, damage caused by improper handling or user alteration, nor unsatisfactory
performance as a result of conditions beyond our control. Lonza Rockland, Inc., shall in no
event be liable for incidental nor consequential damages, including without limitation, lost
profits, loss of income, loss of business opportunities, loss of use and other related damages,
however caused, nor any damage arising from the incorrect use of the product.

License and Trademark Information
Some components and technology of the FlashGel™ System are sold under licensing
agreements. The nucleic acid stain in this product is manufactured and sold under license from
Molecular Probes, Inc. and the FlashGel™ Cassette is sold under license from Invitrogen IP
Holdings, Inc. and is for use only in research applications or quality control, and is covered bypending and issued patents. Dark Reader is a trademark of Clare Chemical Research, Inc. The
FlashGel™ Dock technology contains Clare Chemical Research, Inc. Dark Reader®
transilluminator technology and is covered under US Patents 6,198,107; 6,512,236; and6,914,250. The electrophoresis technology is licensed from Temple University and is covered
under US Patent 6,905,585. Polaroid is a registered trademark of Polaroid Corporation.
RiboPure is a trademark of Ambion, Inc. All other trademarks herein are marks of the Lonza
Group or its affiliates.

For Research Use Only. Not for use in diagnostic procedures.
© 2012 Lonza Rockland, Inc.

37

Lonza Rockland, Inc.
www.lonza.com
scientific.support@lonza.com
Scientific Support: 800-521-0390
Customer Service: 800-638-8174
Document # 00521123-1012-4
Rockland, ME 04841