What is NEB Buffer?
NEB’s restriction enzyme buffer system makes your restriction digests easy and convenient. We are able to offer >210 restriction enzymes that cut in a single buffer, rCutSmart™ . This improves ease-of-use, especially when performing double digests.
What is composition of restriction enzyme buffer?
MULTI-CORE™ Buffer (1X) = 25mM Tris-acetate (pH 7.5 at 37°C), 100mM potassium acetate, 10mM magnesium acetate, 1mM DTT. Notes: 1. For each 10°C rise in temperature between 0°C and 25°C, the pH of Tris buffers decreases 0.31 pH units.
What is BamHI HF?
High-Fidelity (HF®) restriction enzymes have the same specificity as native enzymes, but have been engineered for significantly reduced star activity and performance in a single buffer (rCutSmart™ Buffer). All HF-restriction enzymes come with Gel Loading Dye, Purple (6X).
What does the H in bamh1 mean?
H stands for strain H of Bacillus amyloliquefaciens in BamHI restriction enzyme.
What are NEB enzymes?
NEB has developed a line of High-Fidelity (HF®) restriction enzymes. These engineered enzymes have the same specificity as the native enzyme, with the added benefit of reduced star activity, rapid digestion (5-15 minutes) and 100% activity in CutSmart buffer.
Why BSA is used in restriction digestion?
Adding BSA to a reaction lessens enzyme loss on tube and pipette tip surfaces. BSA stabilizes enzymes in reaction. The stabilizing effects are most pronounced in overnight reactions (Robinson D.
What is Thermo buffer?
Thermo Scientific 10X Buffer O ensures the optimum reaction conditions for restriction enzymes and is premixed with BSA for enhanced stability. Our Five Buffer System ensures the optimum reaction conditions for each restriction enzyme.
What is Tango buffer?
Tango buffer has been designed for double digestions of DNA with conventional restriction enzymes. Visit the Thermo Scientific DoubleDigest tool for more information.
Is BamHI methylation sensitive?
BamHI cannot methylate this modified sequence. Methyltransferase/endonuclease pairs which differ in their sensitivity to non-canonical methylation can be exploited to generate rare DNA cleavage sites.
Does BamHI produce sticky ends?
BamHI binds at the recognition sequence 5′-GGATCC-3′ , and cleaves these sequences just after the 5′-guanine on each strand. This cleavage results in “sticky ends” which are 4 b.p. long. In its unbound form, BamHI displays a central b sheet, which resides in between a helices .
What type of restriction enzyme is BamHI?
BamHI (pronounced “Bam H one”) (from Bacillus amyloliquefaciens) is a type II restriction endonuclease, having the capacity for recognizing short sequences (6 bp) of DNA and specifically cleaving them at a target site.
How do you inactivate BamHI?
Only small amounts of BamHI (up to 10 units) can be inactivated at 80°C in 20 min. To prepare the digested DNA for electrophoresis: – stop the digestion reaction by adding 0.5 M EDTA, pH 8.0 (#R1021), to achieve a 20 mM final concentration. Mix thoroughly, add an electrophoresis loading dye and load onto gel.