Quick Gel Extraction Kit
From CCGB
QIA quick Gel Extraction Kit Protocol
--by Hao Wang
1. Excise the DNA fragment from the agarose gel with a clean, sharp scalpel.
2. Weigh the gel slice in a colorless tube. Add 3 volumes of Buffer QG to 1 volume of gel (100mg~100μl).
3. Incubate at 50 ºC for 10 min (or until the gel slice has completely dissolved). To help dissolve gel, mix by vortexing the tube every 2-3 min during the incubation.
4. After the gel slice has dissolved completely, check that the color of the mixture is yellow.
5. Add 1 gel volume of Isopropanol to the sample and mix. This step increase the yield of DNA fragments <500 bp and >4 kb. For DNA fragments between 500 bp and 4 kb, addition of Isopropanol has no effect on yield. Do not centrifuge the sample at this stage.
6. Place a QIAquick spin column in a provided 2 ml collection tube.
7. To bind DNA, apply the sample to the QIAquick column, and centrifuge for 1 min. The maximum volume of the column reservoir is 800 μl.
8. Discard flow through and place QIAquick column back in the same collection tube.
9. Add 0.75 ml of Buffer PE to QIAquick column and centrifuge for 1 min.
10. Discard the flow-through and centrifuge the QIAquick column for an additional 1 min at 13,000 rpm.
11. Place QIAquick column into a clean 1.5 ml microcentrifuge tube.
12. To elute DNA, add 30-50 μl of Buffer EB (10mM Tris-Cl, pH8.5) or H2O to the center of the QIAquick membrane and centrifuge the column for 1 min.
