Qiagen
QIAEX II Gel Extraction Kit
20021
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QIAEX II Gel extraction kit was used to gel excise and extract DNA inserts of sizes ranging from 15 to 20 kb from 0.7 % agarose gel. Using this kit, I was able to recover 60 % of DNA inserts though with poor quality 260:230 ratio. However, extracted DNA inserts were good enough for downstream cloning.
Extraction of large DNA inserts for restriction cloning.
5 microgram of RE digested DNA having insert size of 15 to 20 kb per well of 0.7 %agarose gel
DNA inserts ranging from 15 to 20 Kb length (each as single band) separated in 0.7 % agarose gel were excised and then extracted using the step-by-step instructions given QIAEX II gel extraction kit. Finally eluted with 50 microliter of nuclease free water. Then quality was assessed by Nanodrop spectrophotometer.
Efficient digestion of agarose gel is must. Use pre-heated (at 70 degree C) nuclease free water for efficient elution/recovery of DNA inserts
Using QIAEX II Gel extraction kit, DNA inserts of size 15 to 20 kb were extracted with good quality (260:280 ratio of 1.8, but 260:230 was 0.4). Recovery was 50 % when compared to input.
N/A
Good for gel extraction of large size DNA inserts from 15 to 25 kb size.
Recovery was not great.
Ideal for extraction and purification large size DNA inserts required for further downstream cloning methods.