Is Gel Loading Dye, Purple (6X) or Gel Loading Dye, Purple (6X), no SDS compatible with other DNA binding dyes such as SYBR® and GelRed™ during gel electrophoresis?.I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA.Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?.How is the improvement in fidelity of HF restriction endonucleases quantitated?.What does it mean to be Time-Saver™ qualified?.Will the HF enzyme produce elevated star activity when used in a buffer other than the one recommended?.
Can the change in buffer preference of the HF enzyme be advantageous?.When should I choose the High Fidelity (HF ®) version of the enzyme?.Why does the HF version of the enzyme have a different recommended buffer than the wild type enzyme?.What does HF® refer to following the name of a restriction enzyme?.What is the difference between NotI-HF and NotI?.Is there a difference in cutting close to the ends between NotI-HF and NotI?.Is there any difference in the methylation sensitivity between NotI-HF and NotI?.coli strain that carries the cloned and modified NotI gene from Nocardia otitidis-caviaru The resolution is at the single nucleotide level.Īn E. The percent degradation is determined by capillary electrophoresis and peak analysis. Restriction Enzyme Competitor Study: Nuclease ContaminationĮcoRI, NotI, and BamHI from multiple suppliers were tested in reactions containing a fluorescent labeled single stranded, double stranded blunt, 3'overhang or 5' overhang containing oligonucleotides. Examples of nuclease contamination studies for some of our HF restriction enzymes are shown below. NEB extensively performs quality controls on all standard and high-fidelity (HF) restriction enzymes. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 with the flexibility to digest overnight without degradation to DNA. HF enzymes also exhibit dramatically reduced star activity. High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer single-buffer simplicity means more straightforward and streamlined sample processing.
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