Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2013/04/22: Difference between revisions
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* Use 125 ng of each primer | * Use 125 ng of each primer | ||
* 10× Reaction Buffer | * 10× Reaction Buffer | ||
100 mM KCl | ** 100 mM KCl | ||
100 mM(NH4)2SO4 | ** 100 mM(NH4)2SO4 | ||
200 mM Tris-HCl (pH 8.8) | ** 200 mM Tris-HCl (pH 8.8) | ||
20 mM MgSO4 | ** 20 mM MgSO4 | ||
1% Triton® X-100 | ** 1% Triton® X-100 | ||
1 mg/ml nuclease-free bovine serum albumin | ** 1 mg/ml nuclease-free bovine serum albumin | ||
(BSA) | (BSA) | ||
* TE Buffer | * TE Buffer | ||
10 mM Tris-HCl (pH 7.5) | ** 10 mM Tris-HCl (pH 7.5) | ||
1 mM EDTA | ** 1 mM EDTA | ||
* '''Control Reaction''' | * '''Control Reaction''' |
Revision as of 06:53, 22 April 2013
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April 19, 2013Quikchange Site Directed Mutagenesis
which includes 5 control reactions.
to multiple freeze-thaw cycles.
(BSA)
5 μl of 10x reaction buffer 2 μl (10 ng) of pWhitescript 4.5-kb control template (5 ng/μl) 1.25 μl (125 ng) of oligonucleotide control primer #1 [34-mer (100 ng/μl)] 1.25 μl (125 ng) of oligonucleotide control primer #2 [34-mer (100 ng/μl)] 1 μl of dNTP mix ddH2O to a final volume of 50 μl
5 μl of 10× reaction buffer X μl (5–50 ng) of dsDNA template X μl (125 ng) of oligonucleotide primer #1 X μl (125 ng) of oligonucleotide primer #2 1 μl of dNTP mix ddH2O to a final volume of 50 μl
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