{"product_id":"neb-r0557s","title":"New England Biolabs, R0557S, BspDI","description":"BspDI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10271509.  \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 rCutSmart™ Buffer Incubate at 37°C 1X rCutSmart™ Buffer 50 mM Potassium Acetate 20 mM Tris-acetate 10 mM Magnesium Acetate 100 µg\/ml Recombinant Albumin (pH 7.9 @ 25°C)  \u003cb\u003eActivity in NEBuffers\u003c\/b\u003e NEBuffer™ r1.1: 25% NEBuffer™ r2.1: 75% NEBuffer™ r3.1: 50% rCutSmart™ Buffer: 100%  \u003cb\u003eDiluent Compatibility\u003c\/b\u003e Diluent A  \u003cb\u003eStorage Buffer\u003c\/b\u003e 10 mM Tris-HCl 50 mM NaCl 1 mM DTT 0.1 mM EDTA 200 µg\/ml Recombinant Albumin 50% Glycerol pH 7.4 @ 25°C  \u003cb\u003eHeat Inactivation\u003c\/b\u003e 80°C for 20 minutes  \u003cb\u003eMethylation Sensitivity\u003c\/b\u003e dam methylation: Blocked by Overlapping dcm methylation: Not Sensitive CpG Methylation: Blocked  \u003cb\u003eFAQ\u003c\/b\u003e Q: Is BspDI  sensitive to methylation? A: Yes it is blocked by overlapping dam methylation, and by CpG methylation at all sites. For up-to-date information about methylation sensitivities, please visit Dam-Dcm and CpG Methylation or REBASE. Q: Does BspDI produce commonly used compatible ends? A: BspDI cleaves to leave a 5´GC which can be ligated to fragments generated by AccI, BstBI, ClaI, TaqI, AciI, BsaHI, HinP1I, HpaII, NarI, Psp1406I. Q: 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. What could be the reason? A: The quality and cleanliness of the DNA are major factors that can affect enzyme activity. Restriction Enzymes active in rCutSmart® Buffer, but not as active in NEBuffer r2.1 and\/or r3.1 (our higher salt buffers), can be inhibited by salt in the reaction. DNA purification procedures that use spin columns can result in DNA solutions with significant levels of salt that can carry over into the reaction and inhibit enzyme activity. To prevent this, we recommend that the DNA solution added to the reaction be no more than 25% of the total reaction volume. This can be attained by adjusting the total reaction volume accordingly. 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":46835487178921,"sku":"R0557S","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-r0557s","provider":"Iright","version":"1.0","type":"link"}