Genenmed® Gel/PCR Kit provides an easy and rapid method for purification of the DNA fragments (100bp to 50Kbp) from the TBE or TAE agarose gels and also from PCR. The entire procedure takes less than 30min and multiple samples can be processed simultaneously. Genenmed® Gel/PCR Kit guarantees higher yield and purity. The purified DNA is ready for various further works, including ligation, labeling, sequencing, cell transfection and restriction digestion without any additional manipulations.
Principle
Genenmed® Gel/PCR Kit utilizes the glass microfiber membrane, to which the DNA fragment of interest binds in the presence of chaotropic salts. The bound DNA is washed and eluted with ddH2O or TE buffer. This simple method eliminates the need for organic solvent extractions and alcohol precipitation.
Kit Contents
Keep all other kit components at room temperature.
Before use, add 52ml absolute ethanol to each bottle of the Buffer GW.
Number of Preps
200
Spin columns
200
Buffer GM
80ml
Buffer GB
120ml
Buffer GW (conc.)
13ml (each bottle)
Chemical Hazard
Both Buffer GM and Buffer GB contains an irritant and can be harmful when contacted with skin, or when inhaled or swallowed. Care should be taken during handling. Always gloves and eye protector, and follow standard safety precautions.
Genenmed® Plasmid Kit Protocol for Gel Extraction
Perform centrifugation at full speed (>10,000Xg or 10,000~13,000rpm)
1. Excise the DNA band of interest from the agarose gel
Do not exceed 3 wells as a processing sample (10ul loading per well)
Cut out the gel slice as thin as possible
2. Transfer the gel slice to a 1.5ml tube and weigh (up to 200mg)
3. Add 400ul Buffer GM
Volume of Baffer GM : weight of gel slice = 2 : 1
4. Incubate at 65~75℃ until the gel slice is completely melted
5. Add 600ul Buffer GB and mix well (pipetting 3~5 times)
6. Transfer the melted mixture to a spin column and centrifuge for 1min
For the DNA fragment (20~50Kbp), centrifuge at 8,000rpm
7. Pour off the flow-through and re-insert the spin column to the collection tube
8. Repeat Step 6,7
9. Apply 600ul Buffer GW and centrifuge for 1min
10. Pour off the flow-through and re-insert the spin column to the collection tube
11. Centrifuge for 3min to completely remove residual Buffer GW
12. Remove spin column from the collection tube and air-dry for 5min
13. Re-insert the spin column to a new 1.5ml tube with no cap
14. Add 30~50ul ddH2O or TE buffer (pH 8.0) and let stand for 1min
15. Centrifuge for 1min
16. Transfer the eluted DNA to a new 1.5ml tube
Genenmed® Plasmid Kit Protocol for PCR Cleanup
Perform centrifugation at full speed (>10,000Xg or 10,000~13,000rpm)
1. Add 250ul Buffer GB to PCR sample (up to 50ul) and mix
Volume of Buffer GB : Volume of PCR sample = 5 : 1
2. Transfer the mixture to a spin column and centrifuge for 1min
3. Pour off the flow-through and re-insert the spin column to the collection tube
4. Apply 600ul Buffer GW and centrifuge for 1min
5. Pour off the flow-through and re-insert the spin column to the collection tube
6. Centrifuge for 3min to completely remove residual Buffer GW
7. Remove spin column from the collection tube and air-dry for 5min
8. Re-insert the spin column to a new 1.5ml tube with no cap
9. Add 50ul ddH2O or TE buffer (pH 8.0) and let stand for 1min
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Tel: 82-2-3474-5800 / Fax: 82-2-3473-4141 / Copyright(C) Genenmed Inc. All Rights Reserved.
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