Gel extractions can be a low yield affair, but overnight incubation over a spin column can increase yields. This protocol details a modified QIAquick Gel Extraction protocol. QIAquick Gel Extraction Kit Cut the desired band out of an agarose gel using a scalpel. Weigh the slice in a snap cap tube and add 3 volumes of QG ...
PCR can be performed directly from a scraped cell colony, using an extended denaturing step to cause lysis of the cells. This is a nice time saving protocol for PCRing targets directly from antibiotic selective plate media. Colony PCR with Patch Plating If subculturing via patch plate, divide a warm LB-Amp plate into a gri...
This protocol details a typical setup for screening for transformed cells in the process of cloning a gene following a typical cloning procedure. An alkaline phosphotase is use to ensure there is no self ligation of the vector. α-Complementation - Many shuttle vectors contain an ampicillin resistance gene so only successfu...
The usefulness of PCR extends beyond replicating a target segment. Unfortunately, PCR cloning can be difficult due to the 3’ adenosine bases added to the ends of PCR products by Taq polymerase. This experiment details cloning a gene from S. cerevisiae such that it is compatible with the TOPO TA cloning process. These adenos...
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