Submit your paper to J Biol Methods today!
0
Videos from Molecular Cloning (View More)
-
PCR Cloning
bioforum - 24 Jul 2013
Videos by bioforum (View More)
-
RNA Isolation and cDNA Synt...
bioforum - 05 Mar 2013
-
How to Pour and Run an Agar...
bioforum - 05 Mar 2013
-
Culture and Passaging of Hu...
bioforum - 05 Mar 2013
-
Chromatin Immunoprecipitati...
bioforum - 05 Mar 2013
-
Microarray Analysis with R
bioforum - 05 Mar 2013
-
Microarrays and Analysis of...
bioforum - 05 Mar 2013
-
Cell Culture Basics from GI...
bioforum - 05 Mar 2013
-
Passaging Cells: GIBCOŽ Cel...
bioforum - 05 Mar 2013
Videos System v3.0.1 © 2021 DevFuse
Couple points. You must use a buffer that contains a detergent such as Taq buffer. Most pfu buffers do not have a detergent and therefor will not efficiently break open the bacterial cells. Also, be aware that false positives are a potential downfall of pcr screening. When you add the ligation mixture to the competent cells during transformation, there is free insert DNA present. That DNA gets spread onto the LB plate and can be picked up by your tip when picking a colony. Always do a second diagnostic test once you have purified the plasmid.