{"product_id":"neb-r0619l","title":"New England Biolabs, R0619L, HpyCH4IV","description":"\u003cb\u003eRelated Categories\u003c\/b\u003e Restriction Endonucleases H M,, 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 pUC19 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: 100% NEBuffer™ r2.1: 50% NEBuffer™ r3.1: 25% rCutSmart™ Buffer: 100%  \u003cb\u003eDiluent Compatibility\u003c\/b\u003e Diluent A  \u003cb\u003eStorage Buffer\u003c\/b\u003e 10 mM Tris-HCl 100 mM NaCl 1 mM DTT 200 µg\/ml BSA 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: Not Sensitive dcm methylation: Not Sensitive CpG Methylation: Blocked  \u003cb\u003eFAQ\u003c\/b\u003e Q: Is HpyCH4IV an isoschizomer of MaeII? A: Yes, it recognizes the same sequence and cleaves at the same place. Both are blocked by SssI (CG) methylase (5-methyl cytosine). Q: Are TaiI and TscI isoschizomers of HpyCH4IV? A: TaiI and TscI are neoschizomers of HpyCH4IV. The recognize the same site but cut at a different location. TaiI is blcoked by SssI and TscI has not been tested. Q: Is HpyCH4IV affected by methylation? A: Yes, it is blocked by 5-methyl cytosine which can be found in mammalian DNA. For up-to-date information about methylation sensitivities, please visit Dam-Dcm and CpG Methylation or REBASE. Q: Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer? A: NEB's Time-Saver™ enzymes have the benefit of working fast (5-15 minutes), but are also designed and qualified to withstand overnight digestions without degradation of DNA. 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":46835521683625,"sku":"R0619L","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-r0619l","provider":"Iright","version":"1.0","type":"link"}