MicroGEM Quick-Start Guide

DNA Extraction Using
forensicGEM Sperm

MicroGEM
Going to extremes for
effortless DNA extraction

Find more information atwww.microgembio.com
or email
info@microgembio.com

MicroGEM International PLC: Research Use Only. All products are subject
to a limited use license. See the product documentation on our website.

QSG_004_190531_forensicGEM SpermforensicGEM
forensicGEM is validated for forensic DNA extractions. Validation
data can be obtained from www.microgembio.com.
General instructions••••

All manipulations should be performed in a clean-room ora PCR hood.
Labcoats, gloves and hairnets should be worn at all times.
Use only certified DNA-free tubes and reagents.
Wash equipment that will come into contact with the sample
in 0.05% bleach. Rinse thoroughly with DNA-free water.

Acrosolv is a mixture of reagents that weaken tissue cell walls. It isdelivered as a lyophilised powder. This should be resuspended inDNA-free water as follows:
Kit size
Code(Rxn)		
50
100
500
1000

FSC0050
FSC0100
FSC0500
FSC1000

Volume ofwater to add
0.55 ml
1.1 ml
5.5 ml11.0 ml

Reagent storage: forensicGEM reagents are stable at room
temperature but on arrival should be stored at 4°C. After tubes
have been opened, the enzyme and the Acrosolv should beplaced at -20°C to safeguard against accidental contamination.
The buffer can remain at 4°C for convenience.
Procedure overview: MicroGEM extraction products use aunique mixture of thermophilic and mesophilic enzymes.
Where a low temperature stage is used, mesophilic cell wall
degrading enzymes come into play. The 75° step then activatesa thermophilic proteinase that lyses the cells, kills nucleases andstrips the DNA of nucleosomes. A final 95° step deactivates thethermophilic proteinase.

SemenStains and Swabs
The processing of the sample will vary dependent on sample type.
For liquid samples, try to keep the volume of the liquid below 10 µl.
With cotton swabs, add 1/4 of the swab directly to the extraction
cocktail. Stained fabric can be swabbed or small portions added
directly.
1. Place or pipette the sample into a thinwalled PCR tube
2. Add:10 µl 10x ORANGE+ Buffer		
2 µl forensicGEM
10 µl AcrosolvWater to 100 µl
3. Mix the sample by vortexing.
4. In a thermal cycler, incubate:						52°C for 5 minutes
75°C for 3 minutes
95°C for 3 minutes

3. Aspirate the extract away from any
residual material.

The DNA is in this solution. Do not discard.
For long term storage of the extracted DNA,
add one tenth volume 10x TE buffer (100 mMTris, pH 7.5, 10 mM EDTA). Store at -20°C.

Technical Tips•

forensicGEM is a preparative method for DNA extraction.
The method lyses cells and removes nucleoproteins fromthe DNA. Extracted DNA can be used for many types ofgenotyping including SNP and STR analysis as well as
quantitative, multiplex and end-point PCR.

•There is no concentration step in the procedure and so theconcentration of the extract is dependent on: 1) The qualityof the sample; 2) In the case of swabs, the type of swab andthe volume of water used to wash the swab; 3) The extraction
volume.

•DNA extracted using forensicGEM is largely single-stranded
because of the 95°C heat step.

•For accurate yield assessment, a qPCR is recommended.
If standard fluorescent chelating dyes are to be used fornormalising samples, then we recommend taking a sampleof the extract before the 95°C step. Alternatively, you cangenerate a standard curve using a previously-made extractthat has been quantified.

•As with any preparative method for nucleic acid extraction,
best results are obtained when samples are handled at 4°C,
or on ice, before and after extraction.

•For long term storage of the extracted DNA, add TE buffer
to 1x (10 mM Tris, pH 7.5, 1 mM EDTA) and store at -20°C.
The forensicGEM reagents are stable at room temperature, butafter tubes have been opened and for longer term storage, theenzymes should be stored at -20°C and the buffers at 4°C.
More information is available at www.microgembio.com