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Cloning problems

Cloning problems - Molecular Biology Techniques

Cloning problems - Molecular Biology Forum. Includes forums for common molecular biology techniques.


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Old 04-10-2012, 09:52 AM
Pipette Filler
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Default Cloning problems



Hi everybody,

I did site-directed mutagensis with pcr (two different pCR product and combine together and amlify template with 5 ans 3 primer containing restriction sites, PCR product size is correct), but I try to clone site-directed pcr product cut by xhoI and spe I ligated into vector pCMV6 cut by same restrion enzymes. I got clone for 3 different mutations but I never get clone for 3 more different mutations.
I also did site-directed on doubl stranded plasmid and ligation, but never get mutated plasmids.

Is there any suggestions for site directed mutagenesis ( any recommendation) and any comment ligation?

Thank you for advance
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