We are doing modifications of an expression vector -a promoter substitution-, and I was wondering if attempting this task through PCR cloning using overlaped homologous sequences could be done by introducing the recombinant product in common strains such as DH5alpha or DH10B (which surely are recA-). If it does, do you know if it is as straightforward as it sounds? Gracias!
Fig. from https://www.ncbi.nlm.nih.gov/pubmed/25359266
You could always use In-fusion cloning instead which does essentially what you are describing.
You can also use the handy primer design tool to help you choose your overlap regions.
http://www.clontech.com/US/Products/Cloning_and_Competent_Cells/Cloning_Resources/Online_In-Fusion_Tools?sitex=10020:22372:US
It comes with the competent cells you would need which have high transformation efficiency, essential for this type of cloning.
http://www.clontech.com/US/Products/Cloning_and_Competent_Cells/Cloning_Kits/Cloning_Kits-HD-Liquid
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