Alm:PCR: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
No edit summary |
No edit summary |
||
| Line 19: | Line 19: | ||
[[Category:Alm]] | [[Category:Alm]] | ||
[[Category: | [[Category:Protocol]] | ||
Latest revision as of 10:58, 3 May 2008
Standard PCR reactions
- ~100 ng template
- ~100 pmole each primer
- buffer to 1X
- polymerase
- denature 94-95°C
- anneal 5°C less than lowest primer hyb temp
- extend 1’/kb to be amplified
PCR Master Mix (2.5X)
- 62.5 U/ml Taq DNA Polymerase
- 125 mM KCl
- 75 mM Tris-HCl, pH 8.3
- 3.75 mM Mg(OAc)2
- 500 uM each dNTP
5'-CATTAG can be useful to add to prevent exonuclease activity from degrading recognition sites
