Ran a PCR to obtain luciferase DNA for sequencing.
Used sea pen gDNA extracted by Jonathan on 20150527.
Master mix calcs are here: 20150702_seapen_PCR
- 95C – 10mins
- 95C – 15s
- 55C – 15s
- 72C – 1min
- Go to Step 2 39 times
Ran samples on 0.8% agarose, low TAE gel stained with EtBr.
Lane 1 – ladder
Lane 2 – empty
Lane 3 – sea pen gDNA
Lane 4 – NTC
PCR did not work. Was expecting a band of ~800bp.
Looks like I may have overloaded the PCR reaction with gDNA. Used 10μL of gDNA.
However, that is quite the smear, suggesting a significant amount of degradation present in the gDNA.
Will re-run this PCR next week with less gDNA (or, cDNA instead) in order to generate a PCR product.