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ligation problem

ligation problem - Molecular Biology Techniques

ligation problem - Molecular Biology Forum. Includes forums for common molecular biology techniques.


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Old 03-24-2013, 09:38 AM
Pipette Filler
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Default ligation problem



Hi. In my gene expression study, I failed to ligate the genes with plasmid PET22b. The genes sizes are 950bp and 1.8kb, respectively. The genes were amplified, excised, RE digested and purified before ligation. The ratio of genes : plasmid are 3:1 in my ligation mixture and the concentration of the gene 1 (950bp) and gene 2 (1.8kb) used are 50 ng and 100 ng, respectively. In the transformation, only the control which is empty plasmid PET22b transformed into E. coli BL21. May I know the possible cause for the failure and how to solve it? Thank you
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