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Gel extraction micro kit

 

1.       Excise the DNA fragment form the agarose gel and weigh it in a 1.5 ml microtube (A gel slice of 100 mg approximately equals to 100 µl). Add 1 volume Buffer GC to the 1.5 ml microtube and incubate the mixture at 55-60oC for 8 min withmixing the tube by tapping the bottom of the tube every 2 min till the gel has melted completely. Cool the tube to RT. For DNA fragment smaller than 200 bp, add 3 volume Buffer GC.
 
2.       Transfer up to 700 µl DNA/Buffer GC mixture to a spin column with a collection tube. Centrifuge at 11,000 x g for 1 min at room temperature. Discard the flowthrough and put the column back to the collection tube. Repeat this step to process the remaining solution.
 
3.       Add 500 µl DNA Wash Buffer to the column and centrifuge at 11,000 x g for 1 min at room temperature. Discard the flow through. Ensure that ethanol has been added to DNA Wash Buffer as instructed before use.
 
4.       Repeat step 3.       
 
5.       Centrifuge the empty column with the lid open at top speed for 1 min to remove the residual ethanol in the column. This is critical for DNA yield.
 
6.       Optional: Spin the empty column at top speed for 1 min.
 
7.       Place the column in a clean 1.5 ml micocentrifuge tube. Add 10-15 µl Elution Buffer or ddH2Oto the column. Incubate at room temperature for 1 min. Centrifuge at 13,000 x g for 1 min to elute the DNA. If it’s a large fragment (≥10 kb), pre-warm the elution buffer to 65oC will increase the yield.
 
7.       Optional: Re-apply the eluate to the column, elute as step 7. This will increase DNA yield.
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