User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/09/05: Difference between revisions
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==September 5, 2013== | ==September 5, 2013== | ||
* '''QuickChange Site Directed Mutagenesis of V0200 to remove | * '''QuickChange Site Directed Mutagenesis of V0200 to remove BSMBI cut site in the Hygro resistance sequence''' | ||
# Use 50 ng of dsDNA template [http://openwetware.org/wiki/User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/05/06 (BD002)] and 125 ng of each primer. | # Use 50 ng of dsDNA template [http://openwetware.org/wiki/User:Behzad_Damadzadeh/Notebook/PcTF_Subcloning_in_E-coli/2013/05/06 (BD002)] and 125 ng of each primer. | ||
# Template strands are about 7kb for BD002. | # Template strands are about 7kb for BD002. | ||
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| Total || 50.0 | | Total || 50.0 | ||
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*No control reaction, I will test the accuracy of the mutagenesis with | *No control reaction, I will test the accuracy of the mutagenesis with BSMBI restriction enzyme. | ||
* Add 1 μl of PfuTurbo DNA polymerase (2.5 U/μl) to the sample reaction | * Add 1 μl of PfuTurbo DNA polymerase (2.5 U/μl) to the sample reaction | ||
* Thermal cycling | * Thermal cycling |
Revision as of 08:56, 13 October 2013
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September 5, 2013
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