Jonesma Lab:Protocols:gelpurification: Difference between revisions
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(New page: <h3><font style="color:#F8B603;">Gel purification</font></h3> ---- #Run DNA out on a gel #Set a waterbath to 50°C #Weigh a 1.5mL tube for each band to be cut #Under a blue light (with an ...) |
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Revision as of 09:56, 8 September 2015
Gel purification
- Run DNA out on a gel
- Set a waterbath to 50°C
- Weigh a 1.5mL tube for each band to be cut
- Under a blue light (with an orange mat as a cutting board), cut out the bands needed (check sizes!) and put into individual tubes
- Reweigh tube with agarose cut out to find weight of agarose
- Add 3x volume of QG buffer (e.g. add 300μL buffer to a 100mg fragment)
- Incubate at 50°C for 10min, mixing occasionally
- Add to blue miniprep column (with clear plastic reservoir attached)- maximum volume 700μL
- Spin at 13 000rpm for 1min
- Empty reservoir and add remainder of dissolved agarose
- Spin at 13 000rpm for 1min
- Add 700μL PE buffer and incubate for 1min
- Spin at 13 000rpm for 1min
- Empty reservoir
- Spin at 13 000rpm for 1min
- Transfer blue column to new 1.5mL tube, labelled appropriately
- Add 40μL dH2O and incubate at room temperature for 1min
- Spin at 13 000rpm for 1min
- Keep tube with DNA, discard used blue column