{"product_id":"supercut™-xhoⅰ-biosci","title":"XhoI-Biosci-SuperCut™","description":"\u003ch2\u003eRestriction Enzyme Site\u003c\/h2\u003e\n\u003cp\u003e5'…C↓T C G A G…3'\u003cbr\u003e3'…G A G C T↑C…5'\u003cbr\u003eIsoschizomers*: PaeR7I, TliI, BssHI, Sfr274I, SlaI, StrI\u003cbr\u003e\u003cspan style=\"color: #00aaff;\"\u003e*Isoschizomers may have different methylation sensitivities.\u003c\/span\u003e\u003c\/p\u003e\n\u003cdiv style=\"text-align: start;\"\u003e\u003cimg src=\"https:\/\/cdn.shopify.com\/s\/files\/1\/0521\/5312\/2997\/files\/20231206151502_160x160.png?v=1701846927\" style=\"margin-bottom: 1px; float: none;\"\u003e\u003c\/div\u003e\n\u003cp style=\"text-align: start;\"\u003e \u003c\/p\u003e\n\u003ch2\u003eStorage Condition\u003c\/h2\u003e\n\u003cp\u003eStorage at -25℃~-15℃ and validity for 24 months.\u003cbr\u003eShipped on Blue Ice.\u003c\/p\u003e\n\u003ch2\u003eComponents\u003c\/h2\u003e\n\u003ctable border=\"1\" style=\"border-collapse: collapse; width: 100%; height: 72px;\"\u003e\n\u003ctbody\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 50%; height: 18px; text-align: center;\"\u003eComponents\u003c\/td\u003e\n\u003ctd style=\"width: 50%; height: 18px; text-align: center;\"\u003eAmount\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 50%; height: 18px; text-align: center;\"\u003eSuperCut™ XhoⅠ (10 units\/μL)\u003c\/td\u003e\n\u003ctd style=\"width: 50%; height: 18px; text-align: center;\"\u003e5000 units\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 50%; height: 18px; text-align: center;\"\u003eSuperCut™ 10×Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 50%; height: 18px; text-align: center;\"\u003e3×1 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 50%; height: 18px; text-align: center;\"\u003e6× Gel Loading Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 50%; height: 18px; text-align: center;\"\u003e1 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003ch2\u003eActivity Definition\u003c\/h2\u003e\n\u003cp\u003eOne unit is defined as the amount of enzyme required to digest 1 μg of λDNA in 50 μL reaction at the optimal reaction temperature in 60 minutes.\u003c\/p\u003e\n\u003ch2\u003eDescription\u003c\/h2\u003e\n\u003cp\u003eSuperCut™ Restriciton Enzymes are a series of genetically engineered restriction enzymes that can accurately complete DNA digestion in 5~15 minutes, suitable for rapid digestion of plasmid DNA, PCR products or genomic DNA.\u003cbr\u003eSuperCut™ Restriciton Enzymes has the following characteris-tics: digestion can be completed within 5~15 minutes; share a diges-tion buffer, which greatly simplifies the digestion reaction system; good enzyme activity redundancy for easy substrate overload or com-plex template digestion. In addition, dephosphorylation and ligation reagent of Biosci TM is 100% active in SuperCut™ Buffer, supporting one-tube reaction and improving the experience of \"digestion-ligation- redigestion\".\u003c\/p\u003e\n\u003ch2\u003eRecommended Reaction Conditions\u003c\/h2\u003e\n\u003cp\u003e1× SuperCut™ Buffer；\u003cbr\u003eIncubate at 37℃；\u003cbr\u003eRefer to \"Protocol for Fast DNA Digestion\" for reaction setup.\u003c\/p\u003e\n\u003ch2\u003eHeat Inactivation\u003c\/h2\u003e\n\u003cp\u003e1.Incubation at 80℃ for 20 minutes.\u003cbr\u003e2.Add an appropriate amount of 6 × Gel Loading Buffer according to the reaction system to terminate the reaction.\u003c\/p\u003e\n\u003ch2\u003eQuality Control\u003c\/h2\u003e\n\u003cp\u003e\u003cstrong\u003eFunctional Test\u003c\/strong\u003e\u003cbr\u003eA 20 μL reaction in SuperCut™ Buffer containing 1 μg of λDNA (HindIII digest) and 10 units of SuperCut™ XhoⅠ incubated for 15 minutes at 37℃ results in complete digestion as determined by agarose gel electrophoresis.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eProlonged Incubation \/ Star Activity Assay\u003c\/strong\u003e\u003cbr\u003eA 20 μL reaction in SuperCut™ Buffer containing 1 μg of λDNA (HindIII digest) and 10 units of SuperCut™ XhoⅠ incubated for 3 hours at 37℃ results in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis. Longer incubation may result in star activity.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eEnzyme Digestion-Ligation-Redigestion Test\u003c\/strong\u003e\u003cbr\u003eAt the optimal reaction temperature, the DNA was digested using 10 units of SuperCut™ XhoⅠ, and then the digestion product was recovered. The DNA fragments can be religated using an appropriate amount of T4 DNA Ligase at 22℃. After the ligation product is recovered again, the ligation product can be recut using SuperCut™XhoⅠ.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eNon-specific Endonuclease Activity\u003c\/strong\u003e\u003cbr\u003eAt the optimal reaction temperature, 10 units of SuperCut™ AvrII was incubated with a 20 μL reaction in SuperCut™ Buffer containing 1 μg of supercoiled plasmid DNA for 4 h, and the plasmid DNA was still supercoiled detected using agarose gel electrophoresis.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eBlue\/White Screening Assay\u003c\/strong\u003e\u003cbr\u003eAn appropriate vector containing lacZα gene is digested by 10 units of SuperCut™ XhoⅠ. The digested product is ligated and transformed into E.coli competent cell. On Luria- Bertani culture plate with X-Gal, IPTG and appropriate antibiotic, the successfully ligated β-galactosidase gene can be\u003cbr\u003eexpressed and gives rise to a blue colony, while an interrupted gene (i.e. degraded DNA end) gives rise to a white colony. SuperCut™ restriction enzymes must produce fewer than 1% white colonies.\u003c\/p\u003e\n\u003ch2\u003eMethod of application\u003c\/h2\u003e\n\u003cp\u003e\u003cstrong\u003e1. Protocol for Fast DNA Digestion\u003c\/strong\u003e\u003cbr\u003e① Combine the following reaction components on ice in the order indicated：\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse: collapse; width: 100%; height: 90px;\"\u003e\n\u003ctbody\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 34.8394%; height: 18px; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 21.4263%; height: 18px; text-align: center;\"\u003ePlasmid DNA\u003c\/td\u003e\n\u003ctd style=\"width: 20.7962%; height: 18px; text-align: center;\"\u003ePCR product\u003c\/td\u003e\n\u003ctd style=\"width: 22.9381%; height: 18px; text-align: center;\"\u003eGenomic DNA\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 34.8394%; height: 18px; text-align: center;\"\u003eDNA\u003c\/td\u003e\n\u003ctd style=\"width: 21.4263%; height: 18px; text-align: center;\"\u003e≤ 1 μg\u003c\/td\u003e\n\u003ctd style=\"width: 20.7962%; height: 18px; text-align: center;\"\u003e≤0.2 μg\u003c\/td\u003e\n\u003ctd style=\"width: 22.9381%; height: 18px; text-align: center;\"\u003e≤ 5 μg\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 34.8394%; height: 18px; text-align: center;\"\u003eSuperCut™ XhoⅠ\u003c\/td\u003e\n\u003ctd style=\"width: 21.4263%; height: 18px; text-align: center;\"\u003e10 units\u003c\/td\u003e\n\u003ctd style=\"width: 20.7962%; height: 18px; text-align: center;\"\u003e10 units\u003c\/td\u003e\n\u003ctd style=\"width: 22.9381%; height: 18px; text-align: center;\"\u003e30-50 units\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 34.8394%; height: 18px; text-align: center;\"\u003eSuperCut™ 10×Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 21.4263%; height: 18px; text-align: center;\"\u003e2 μL\u003c\/td\u003e\n\u003ctd style=\"width: 20.7962%; height: 18px; text-align: center;\"\u003e3 μL\u003c\/td\u003e\n\u003ctd style=\"width: 22.9381%; height: 18px; text-align: center;\"\u003e5 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 34.8394%; height: 18px; text-align: center;\"\u003eddH 2 O\u003c\/td\u003e\n\u003ctd style=\"width: 21.4263%; height: 18px; text-align: center;\"\u003eTo 20 μL\u003c\/td\u003e\n\u003ctd style=\"width: 20.7962%; height: 18px; text-align: center;\"\u003eTo 30 μL\u003c\/td\u003e\n\u003ctd style=\"width: 22.9381%; height: 18px; text-align: center;\"\u003eTo 50 μL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003ea. DNA should be free of phenol, chloroform, ethanol, EDTA, detergents or high concentrations of salts, otherwise enzyme activity will be affected; Methylated DNA inhibits certain restriction enzyme digestion reactions.\u003c\/p\u003e\n\u003cp\u003e② Gently suck or flick the tube wall to mix well (never vortex), then centrifuge instantaneously to collect reaction solution；\u003cbr\u003e③ Incubate at 37℃ for 15 minutes (plasmid DNA) or for 15~30 minutes (PCR product) or for 30~60 minutes (genomic DNA)；\u003cbr\u003e④ Optional: Inactivate the enzyme by heating for 20 minutes at 80℃ or add an appropriate amount of 6× Gel Loading Buffer according to the reaction system to terminate the reaction.\u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003e2. Double and Multiple Digestion of DNA\u003c\/strong\u003e\u003cbr\u003e① Use 10 units of each enzyme and scale up the reaction conditions appropriately；\u003cbr\u003e② The combined volume of the enzymes in the reaction mixture should not exceed 1\/10 of the total reaction volume；\u003cbr\u003e③ If the enzymes require different reaction temperatures, start with the enzyme that requires a lower temperature, then add the second enzyme and incubate at the higher temperature.\u003cbr\u003e\u003cspan style=\"color: #00aaff;\"\u003eNote: If the total volume of reaction solution is larger than 20 μL, the incubation time should be increased appropriately, and water bath, metal bath, or sand bath should be used as much as possible.\u003c\/span\u003e\u003c\/p\u003e\n\u003cp\u003e\u003cspan style=\"color: #00aaff;\"\u003e\u003cstrong\u003e\u003cspan style=\"color: #000000;\"\u003eNumber of Recognition Sites in DNA\u003c\/span\u003e\u003c\/strong\u003e\u003cbr\u003e\u003c\/span\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 100%;\" border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003eλDNA\u003c\/td\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003eΦX174\u003c\/td\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003epBR322\u003c\/td\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003epUC57\u003c\/td\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003epUC18\/19\u003c\/td\u003e\n\u003ctd style=\"width: 11.6163%; text-align: center;\"\u003eSV40\u003c\/td\u003e\n\u003ctd style=\"width: 13.9728%; text-align: center;\"\u003eM13mp18\/19\u003c\/td\u003e\n\u003ctd style=\"width: 11.9109%; text-align: center;\"\u003eAdeno2\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 12.5%; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 11.6163%; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 13.9728%; text-align: center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"width: 11.9109%; text-align: center;\"\u003e6\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethylation Effects on Digestion\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable style=\"border-collapse: collapse; width: 100%; height: 36px;\" border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eDam\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eDcm\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eCpG\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eEcoKI\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eEcoBI\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eNo effect\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eNo effect\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eimpaired\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eNo effect\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eNo effect\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e \u003c\/p\u003e\n\u003cp\u003e\u003cstrong\u003eActivity in Different Buffers\u003c\/strong\u003e\u003c\/p\u003e\n\u003ctable border=\"1\" style=\"border-collapse: collapse; width: 100%; height: 36px;\"\u003e\n\u003ctbody\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 16.7599%; height: 18px; text-align: center;\"\u003e \u003c\/td\u003e\n\u003ctd style=\"width: 23.2401%; height: 18px; text-align: center;\"\u003eSuperCut™\u003cbr\u003eBuffer\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eThermo Scientific\u003cbr\u003eFastDigest Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eNEB\u003cbr\u003eCutSmart ® Buffer\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003eTakara\u003cbr\u003eQuickCut™ Buffer\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr style=\"height: 18px;\"\u003e\n\u003ctd style=\"width: 16.7599%; height: 18px; text-align: center;\"\u003eActivity\u003c\/td\u003e\n\u003ctd style=\"width: 23.2401%; height: 18px; text-align: center;\"\u003e100%\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003e100%\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003e100%\u003c\/td\u003e\n\u003ctd style=\"width: 20%; height: 18px; text-align: center;\"\u003e100%\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003cp\u003e\u003cspan style=\"color: #00aaff;\"\u003eNote: The activity data is from the standard reation test of Biosci\u003cspan\u003e™\u003c\/span\u003e Restriction Enzyme described above.\u003c\/span\u003e\u003cbr\u003e\u003c\/p\u003e","brand":"Dakewe","offers":[{"title":"5000 units","offer_id":43541335736501,"sku":"8052441","price":65.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0521\/5312\/2997\/files\/8052441SuperCut_XhoIFastRestrictionEndonucleasespng.png?v=1732886496","url":"https:\/\/biofargo.com\/products\/supercut%e2%84%a2-xho%e2%85%a0-biosci","provider":"Biofargo","version":"1.0","type":"link"}