Restriction

... eventually break down. T4 ligase works just fine at RT, but you can't reuse it now, can you? In fact, that is the main paradigm with enzymes. take restriction enzymes: most of them are active at 37C, but they need to be stored at -20C, and even when you take them out of the freezer you should keep ...

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... much lost." My question is: Ligase has 100% activity at room temperature 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 ...

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... 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 the fragments using Qiagen kit. Using the same enzymes (Kpn1 and XbaI) restriction ...

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... 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 the fragments using Qiagen kit. Using the same enzymes (Kpn1 and XbaI) restriction ...

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... are all thru out the mammalian cell cycle: Mid-G1 cyclin CDKs - Cyclin D-CDK4, Cyclin D-CDK6 (complexes form in mid G1 phase and peak at the restriction point, then taper off at S phase) Late-G1 cyclin CDK - Cyclin E-CDK2 (complexes starts right after the restriction point, peaks at the beginning ...

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