927
edits
Project:Blood typing: Difference between revisions
From London Hackspace Wiki
→Extraction and PCR (Simon, Lui, Will) 26/2/13
Mycoplasma (talk | contribs) No edit summary |
Mycoplasma (talk | contribs) |
||
| Line 663: | Line 663: | ||
* Did two reactions for each gDNA sample. 25ul total PCR reaction volume - 5ul dH20, 10ul template, 2.5ul each forward & reverse primers, 12.5ul Taq readymix | * Did two reactions for each gDNA sample. 25ul total PCR reaction volume - 5ul dH20, 10ul template, 2.5ul each forward & reverse primers, 12.5ul Taq readymix | ||
* Initial D - 96C 5 mins, then 40 cycles of D-96C 1 min, A-55C 30 secs, E-72C 1 min. | * Initial D - 96C 5 mins, then 40 cycles of D-96C 1 min, A-55C 30 secs, E-72C 1 min. | ||
* Ran samples on gel to check PCR succeeded. | * Ran samples on gel to check PCR succeeded. JJ's did, couldn't tell with Bene's as we fucked up loading it onto the gel. | ||
===Restriction digest=== | ===Restriction digest=== | ||
* Incubated JJ's PCR product with Kpn1 at 37C for 65 mins. Restriction digest mix contained 25ul PCR product, 1ul Kpn1, 15ul buffer. | * Incubated JJ's PCR product with Kpn1 at 37C for 65 mins. Restriction digest mix contained 25ul PCR product, 1ul Kpn1, 15ul buffer. | ||
* Ran result on a gel. PCR product remained undigested. | |||
