{"product_id":"neb-r0547l","title":"New England Biolabs, R0547L, PacI","description":"PacI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10128529.  \u003cb\u003eRelated Categories\u003c\/b\u003e Restriction Endonucleases P R,, 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 pNEB193 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: 75% NEBuffer™ r3.1: 10% rCutSmart™ Buffer: 100%  \u003cb\u003eDiluent Compatibility\u003c\/b\u003e Diluent A  \u003cb\u003eStorage Buffer\u003c\/b\u003e 10 mM Tris-HCl 200 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 65°C for 20 minutes  \u003cb\u003eMethylation Sensitivity\u003c\/b\u003e dam methylation: Not Sensitive dcm methylation: Not Sensitive CpG Methylation: Not Sensitive  \u003cb\u003eFAQ\u003c\/b\u003e Q: How can the efficieny of PacI be increased? A: Incubation overnight at 4°C followed by a 2 hour incubation at 37°C works much better than 37°C overnight for genomic DNA embedded in agarose. Q: Is PacI inhibited by salt? A: PacI is inhibited by salt concentrations over 100mM. Q: What is the activity of PacI at 25°C? A: PacI is 50% active at 25°C. 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: Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)? A: All HF-restriction enzymes and most NEB non-HF restriction enzymes are supplied with Gel Loading Dye, Purple (6X). The non-HF restriction enzymes that come supplied with purple dye are: AatII AsiSI BsmBI-v2 EagI MboI NlaIII PvuI SmaI Acil AvrII BspHI EcoRI MboII NotI RsaI SpeI AfeI BamHI BsrGI EcoRV MluI NspI SacI StuI AflII BbsI ClaI Esp3I FseI MseI PacI SacII AgeI BglII DdeI HaeIII MspI PciI SalI XbaI AluI BsaI DpnI HindIII NcoI PmeI SapI XhoI ApaI BseYI DpnII HpaI NdeI PsiI-v2 SfaNI XmaI ApeKI BsiWI DraI KpnI NheI PstI SfiI XmnI AscI * All HF restriction enzymes are also supplied with Gel Loading Dye, Purple (6X) Q: Is this enzyme sensitive to dam, dcm or mammalian CpG methylation? A: No. This enzyme is not sensitive to dam, dcm, or mammalian CpG methylation. For up-to-date information about methylation sensitivities, please visit Dam-Dcm and CpG Methylation or REBASE. Q: Does NEB offer any BSA-free and\/or animal origin-free restriction enzymes for linearization of plasmids for mRNA vaccine development? A: Yes, NEB has formulated some restriction enzymes (including BspQI, an SapI isoschizomer) without animal-derived components and no BSA appears in their final formulation. In addition, we have several restriction enzymes available without BSA nor other animal-derived components in their final formulation. This list includes: BbsI-HF BsaI-HFv2 BspQI, isoschizomer of LguI, GMP-grade* now available BspQI-HF ClaI HindIII-HF PacI SapI SpeI SwaI XbaI XhoI XmnI Other enzymes can be formulated upon request. Please inquire here. * “GMP Grade” and “GMP-grade” are branding terms NEB uses to describe products manufactured or finished at NEB’s Rowley facility. The Rowley facility was designed to manufacture products under more rigorous infrastructure and process controls to achieve more stringent product specifications and customer requirements. Products manufactured at NEB’s Rowley facility are manufactured in compliance with ISO 9001 and ISO 13485 quality management system standards. However, at this time, NEB does not manufacture or sell products known as Active Pharmaceutical Ingredients (APIs), nor does NEB manufacture its products in compliance with all of the Current Good Manufacturing Practice regulations. 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. Q: Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures? A: The recommended storage temperature for Gel Loading Dye, Purple 6X, is room temperature (25°C). Commonly, the SDS may precipitate out of solution when the dye is stored on cold temperatures. This slight precipitate can be dissipated by letting the vial sit at room temperature for an hour, or warming gently for 10 minutes. Mix the vial a few times before use, to bring the solution back to homogeneity. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835506380969,"sku":"R0547L","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-r0547l","provider":"Iright","version":"1.0","type":"link"}