DNA Extraction and PCRs on Passage 4 DiNV Used to Inoculate Cells
Rob wanted me to make sure that the virus solution that I gave the cells that I used for DNA extraction for electroporation worked on all the PCR primers I use. So I combined 2 tubes of passage 4 DiNV aliquots for DNA extraction, then ran p47, 115, lef 4, and lef 9 PCRs on it.
DNA Extraction followed exactly the general protocol except that DNA was only resuspended for 30min before use in PCRs
4 PCRs were run on the samples, and the process followed the general PCR protocol completely. Master mix volumes are listed here:
| reagent | p47 | 115 | lef 9 | lef 4 |
|---|---|---|---|---|
| GoTaq | 17.5ul | 17.5ul | 17.5ul | 17.5ul |
| F primer | 0.875ul | 0.875ul | 0.875ul | 0.875ul |
| R primer | 0.875ul | 0.875ul | 0.875ul | 0.875ul |
| molecular grade water | 12.25ul | 12.25ul | 12.25ul | 12.25ul |
All PCR programs were run for 35 cycles and program information can be found here.
A 1% gel was run at 90V for 45 minutes to resolve the bands:
