Gel Extraction (Qiagen) |
· Excise the DNA fragment from the Agarose gel with a clean, sharp scalpel. (Minimize the size of the gel by removing extra agarose).
· Weigh the gel slice in a colourless tube. Add 3 volumes of Buffer QG to 1 volume of gel (100 mg~300µl).
· Incubate at 50°C for 10 mins. to dissolve agarose. Vortexing in between helps (37°C is fine).
· Add 1 gel volume of iso-propanol to the sample and mix (not for 500bp-4kb).
· Place a QIAquick spin column in a provided 2 ml collection tube.
· To bind DNA, apply the sample to the QIAquick column and centrifuge for 1min (maximum volume is 800µl, so do twice).
· Discard flow-through and place QIAquick column back in the same collection tube.
· Add 750µl Buffer PE to the column and centrifuge for 1min.
· Discard flow-through and centrifuge the column for an additional 1 min. at 13,000 rpm.
· Place the column into a clean 1.5ml eppendorf.
· To elute DNA, add 30µl Buffer EB or H2O to the centre of the QIA membrane and centrifuge for 1 min.
· Can store at -20°C.
|
Tuesday, 30 November 2010 10:53 |