User:Stuart McKellar/Notebook/Bird Sex Testing/2013/01/19

From OpenWetWare
Jump to navigationJump to search
The printable version is no longer supported and may have rendering errors. Please update your browser bookmarks and please use the default browser print function instead.
Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Preliminary results generation

  • I am getting results that Iam confident are usable. This is by comparing them to the positive control samples and getting ratios that are correct.

I will be doing a PCR with P2/P8 primers and then comparing against positive controls with ALL SAMPLES (except loris). Then I will repeat this with the 2550F primer pair at 55°C and comparing all these results. If I can't get consistency, I will order the NP MP primer set.

Method:

black pen - P2 - 60C blue - 2550F - 55C

29 samples


30 samples master mix

Tube

  • 1ul F primer
  • 1ul R primer
  • 2.0ul dNTPs
  • 0.5u Taq
  • 1ul DNA
  • 2.5ul 10x buffer
  • 1.5ul MgCl2
  • 16ul H2O


Master mix

  • 30ul F primer yes
  • 30ul R primer yes
  • 60ul dNTPs yes
  • 3ul Taq (0.5u per tube)
  • 75ul 10x buffer
  • 45ul MgCl2
  • 480ul H2O

added 35ul oil

3min initial melt\

2550 sample

Master mix

  • 30ul F primer yes
  • 30ul R primer yes
  • 60ul dNTPs yes
  • 3ul Taq (0.5u per tube)
  • 75ul 10x buffer yes
  • 45ul MgCl2 yes
  • 480ul H2O yes


Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Stuart_McKellar/Notebook/Bird_Sex_Testing/2013/01/19&oldid=668851"