See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes. ZERO BIAS - scores, article reviews, protocol conditions and more BsmBI This product has been replaced with an updated version engineered for improved Golden Gate Assembly, BsmBI-v2 (NEB #R0739 ).
Because we are cloning an ORF, we want to clone from the start codon (ATG) to the stop codon (TGA, in this example).
We first randomly extracted 9824 target sequences from the GeCKOv1 library , excluding those with BsmBI sites in their sequences. 240 County Road Ipswich, MA 01938-2723 978-927-5054 (Toll Free) 1-800-632-5227 Fax: 978-921-1350 [email protected] Assuming you are amplifying from plasmid DNA (rather than from genomic DNA or a cDNA library), roughly 18-21bp is usually sufficient to give specificity and to … I'm assuming you digested your PCR product with BsmBI, gel purified and are cloning that into pDNR221 that you've cut with BsmBI (is that correct?). BsmBI-v2 is an engineered and improved version of BsmBI. Ensure you are using only one Type IIS restriction enzyme, not two distinct ones (one of BsaI will work; one of BsaI and one of BsmBI will not work) Then, check that your insertion region is correct: 1. Catalog # R0580 was discontinued on March 31, 2020 ; An E. coli strain that carries the modified cloned BsmBI gene from Bacillus stearothermophilus B61. Recognition sequence, reaction conditions, heat denaturation, and microbial source for BsmBI restriction enzyme. Locate commercially available restriction enzymes by category, name, recognition sequence, or overhang.
(NEB uses the designation "v2" or "HF" in the name to indicate engineered enzymes.) RNAi, Oligos, Assays, Gene Editing & Gene Synthesis Tools Oligos Tools. From 1841 target sequences from the coding sequences of three human and six mouse cell surface marker–encoding genes ... and the BsmBI cut sites … Note: Also available as a FastDigest enzyme for rapid DNA digestion. Every barcode and BsmBI cut site are unique to a given subpool minimizing the chance for undesired assembly. It's more likely that you had unligated cut … Ec, Eco P15I; Ac, Acu I. I don't actually think you can.
Please indicate how you would like the restriction sites displayed Map of restriction sites Table of sites, sorted alphabetically by enzyme name Table of sites, sorted sequentially by base pair number. Eurofins MWG Operon Oligos Tool Poly(A), polyadenylate.
Bioz Stars score: 93/100, based on 175 PubMed citations. Cut at CGTCTC(1/5)^ sites with Thermo Scientific™ FastDigest™ BsmBI (Esp3I), which peforms best at 37°C in 5-15 minutes using universal FastDigest Buffer. Please indicate which enzymes to include in the display All enzymes Enzymes not cutting Find out where the cut-sites are located on the plasmid (using the restriction digest tool) and highlight in … ( B ) Type III and type IIS restriction sites to cut out the 20-bp guide sequence. For instance if you have a vector that has two asymmetric BsmBI cut sites, then phosphatase treatment will prevent re-ligation of vectors that have only been cut … Bsmbi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 25 PubMed ... ( A ) Semi-random primer. Find sites which may be introduced by silent mutagenesis. Thermo Scientific Esp3I (BsmBI) restriction enzyme recognizes CGTCTC(1/5)^ sites and cuts best at 37°C in Tango (+DTT) buffer. The genetic handle is designed with outward-facing BsaI recognition sites that enable cutting within the beginning and ends of short flexible serine–glycine linkers.