{"product_id":"neb-r0545s","title":"New England Biolabs, R0545S, BcgI","description":"BcgI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10271508.  \u003cb\u003eRelated Categories\u003c\/b\u003e Restriction Endonucleases B  \u003cb\u003eApplications\u003c\/b\u003e Restriction Enzyme Digestion  \u003cb\u003eSpecification\u003c\/b\u003e \u003cb\u003eUnit Definition\u003c\/b\u003e One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 37°C in a total reaction volume of 50 μl.  \u003cb\u003eReaction Conditions\u003c\/b\u003e 1X NEBuffer™ r3.1 Incubate at 37°C 1X NEBuffer™ r3.1 100 mM NaCl 50 mM Tris-HCl 10 mM MgCl2 100 µg\/ml Recombinant Albumin (pH 7.9 @ 25°C)  \u003cb\u003eActivity in NEBuffers\u003c\/b\u003e NEBuffer™ r1.1: 10% NEBuffer™ r2.1: 75% NEBuffer™ r3.1: 100% rCutSmart™ Buffer: 50%  \u003cb\u003eDiluent Compatibility\u003c\/b\u003e Diluent A  \u003cb\u003eStorage Buffer\u003c\/b\u003e 10 mM Tris-HCl 50 mM KCl 1 mM DTT 0.1 mM EDTA 200 µg\/ml Recombinant Albumin 0.32 mM S-adenosylmethionine (SAM) 50% Glycerol pH 7.4 @ 25°C  \u003cb\u003eHeat Inactivation\u003c\/b\u003e 65°C for 20 minutes  \u003cb\u003eMethylation Sensitivity\u003c\/b\u003e dam methylation: Impaired by Overlapping dcm methylation: Not Sensitive CpG Methylation: Blocked by Some Combinations of Overlapping  \u003cb\u003eFAQ\u003c\/b\u003e Q: Does BsgI require SAM for activity? A: Yes, S-adenosylmethionine (SAM) is required for optimal activity (SAM is supplied in the enzyme formulation as of October 2020). If you are unsure whether SAM is in the enzyme formulation and you have a vial of SAM from NEB, it is fine to add SAM to the reaction per previous guidelines. Q: Do BcgI generated ends ligate? A: BcgI excises a 32 bp fragment with 2-base 3´ overhangs. These overhangs are not specific, so whether the ends can be ligated depends entirely on the specific DNA sequence being cut. Q: Is this enzyme sensitive to dam, dcm or mammalian CpG methylation? A: Yes. This enzyme cannot cut recognition sites that are impaired by overlapping dam methylation or blocked by some combinations of overlapping CpG methylation. For up-to-date information about methylation sensitivities, please visit Dam-Dcm and CpG Methylation or REBASE. Q: Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers? A: NEB is excited to announce that we have switched BSA-containing reaction buffers (NEBuffer 1.1, 2.1, 3.1 and CutSmart® Buffer) to Recombinant Albumin-containing buffers (NEBuffer r1.1, r2.1, r3.1 and rCutSmart™ Buffer). We are also in the process of moving all restriction enzyme formulations to contain rAlbumin. We feel that moving away from animal-containing products is a step in the right direction and are able to offer this enhancement at the same price. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835535151273,"sku":"R0545S","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-r0545s","provider":"Iright","version":"1.0","type":"link"}