{"product_id":"neb-r0149s","title":"New England Biolabs, R0149S, TaqI-v2","description":"  \u003cb\u003eRelated Categories\u003c\/b\u003e Restriction Endonucleases T Z,, Time-Saver Qualified Restriction Enzymes  \u003cb\u003eApplications\u003c\/b\u003e Fast Cloning: Accelerate your cloning workflows with reagents from NEB,, 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 65°C in a total reaction volume of 50 μl.  \u003cb\u003eReaction Conditions\u003c\/b\u003e 1X rCutSmart™ Buffer Incubate at 65°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: 50% NEBuffer™ r2.1: 100% NEBuffer™ r3.1: 50% rCutSmart™ Buffer: 100%  \u003cb\u003eDiluent Compatibility\u003c\/b\u003e Diluent B  \u003cb\u003eStorage Buffer\u003c\/b\u003e 10 mM Tris-HCl 300 mM NaCl 1 mM DTT 0.1 mM EDTA 500 µg\/ml Recombinant Albumin 50% Glycerol pH 7.4 @ 25°C  \u003cb\u003eHeat Inactivation\u003c\/b\u003e No  \u003cb\u003eMethylation Sensitivity\u003c\/b\u003e dam methylation: Blocked by Overlapping dcm methylation: Not Sensitive CpG Methylation: Not Sensitive  \u003cb\u003eActivity at Temperature\u003c\/b\u003e @37°C: 10%  \u003cb\u003eFAQ\u003c\/b\u003e Q: Why isn't TaqI-v2 cutting? A: 1. It could be inhibited by some mini-prep impurities. Drop-dialyse or ethanol wash. 2. It is blocked by overlapping dam methylation. 3. PCR products can be resistant to cleavage. Purify the DNA. 4. Incubation without BSA results in 50% activity. 5. Incubation at 37°C results in 10% activity. Q: Does a certain pH range inhibit TaqI-v2  activity? A: Yes, activity is decreased if the reaction buffer pH is not between 7.5 and 8.5. Q: Does TaqI-v2 recognize mutliple sites? A: The enzyme can recognise A\/C or G\/T mismatches (NAR 14: 459-6590: 1943-1949 (1986) Jiricny, J. Martin, D. ). The star sites are a single base pair difference (Gene 65: 149-165, (1988) Barany, F.). Q: Is this enzyme sensitive to dam, dcm or mammalian CpG methylation? A: Yes. This enzyme cannot cut recognition sites that are blocked by overlapping dam 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":46835513884841,"sku":"R0149S","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-r0149s","provider":"Iright","version":"1.0","type":"link"}