Shefigem2010: Gel extraction

Gel Extraction

This method was taken from the QIAGEN hand book provided with the Gel extraction kit:

1.) Excise the DNA fragment from the agarose gel with a clean, sharp scalpel.

2.) Weigh the gel slice. Add 3 volumes of Buffer QG to 1 volume of gel. ( e.g. add 300μL of buffer to every 100mg of gel)

3.) Incubate at 50°C and mix until gel has completely dissolved.

4.) Add 1 gel volume of isopropanol to the sample and mix. (e.g. add 100μL of isopropanol to every 100mg of gel)

5.) Pour or pipette the sample into a QIAquick spin column provided with the kit and centrifuge for 1min at 13,000rpm.

6.) Discard flow-through.

7.) Add 0.5ml of Buffer QG to the spin column and centrifuge for 1min at 13,000rpm, and discard flow through.

8.) To wash, add 0.75ml of Buffer PE to the spin column and centrifuge for 1min at 13,000rpm, and discard flow through.

9.) Centrifuge the sample for an additional 1min at 13,000rpm.

10.) Place the column into a clean 1.5ml microcentrifuge tube.

11.) To elute DNA, add 50μL of distilled, autoclaved water to the centre of the column and centrifuge for 1min at 13,000rpm.