Expression vector

... and even when you take them out of the freezer you should keep them on ice or even better use an enzyme cooler. 2) Theoretically you're right: the vector shouldn't re-ligate. Unfortunately, in reality it does re-ligate, and sometimes with quite good efficiency (the reason is mainly that bases can ...

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... then how can it go bad when left at RT? 2)Since I used two different restriction enzymes producing overhangs of different bases how can the vector religate, in other words how would the CIP help, wouldnt using CIP make more sense in blunt end producing enzymes? I had followed points 4 and ...

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Hello, I have two problems: 1) We ordered pOTB7 vector with gene A from a company and constructed deletion constructs of the gene A from above vector. We designed primers with restriction sites for 4 deletion constructs, PCR amplified and gel purified ...

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Hello, I have two problems: 1) We ordered pOTB7 vector with gene A from a company and constructed deletion constructs of the gene ... at 5 different occasions and no positive clones were obtained. SInce the expression of the insert has been frustating and unsucessful we decided to cut ...

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... call it GENE for the sake of clarity) into the MCS of the pEGFP-N1 vector from Clontech. The GENE is followed by a poly A signal, the whole ... of the mRNA into the EGFP coding sequence and, thus, prevent the expression of EGFP in cells following transfection. After Cre-mediated recombination, ...

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