Toronto/Lab Protocols/Extraction


Gel Extraction

3 hours including ligation

Place the gel on the transilluminator to view the stained DNA. PLEASE make sure the UV-shield is down before turning on the UV. Check to see if there are the expected number of bands, and that the bands are at the expected lengths.

In order to obtain the desired DNA strand, you just physically remove it from the gel. PLEASE DO NOT CUT DIRECTLY ON THE TRANSILLUMINATOR, USE THE PLASTIC SHEET. In order to extract the gel, using a blade, carefully cut out the desired band. Try to cut as close to the DNA band as possible. It is important that the mass of the gel is as low as possible (<140 mg is a good rule of thumb) because the final yield of DNA, after the purification process, decreases as gel volume increases. Gel Purification

  1. For every gel extract, heat up 30 μL of TE Buffer to 70 °C.
  2. Follow the instructions provided with the Invitrogen Gel Purification Kit. Except, at the last step, to further concentrate the DNA, only add 30 μL of the warm TE Buffer to each tube.

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