{"product_id":"biofargo-bsai-gmp-grade","title":"Biofargo BsaI, GMP Grade","description":"\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eDescription\u003c\/h2\u003e\n\u003cp\u003eBiofargo BsaI, GMP Grade is a restriction endonuclease produced by recombinant expression in \u003cem\u003eE. coli\u003c\/em\u003e and manufactured under GMP specifications, for use as a raw material in mRNA, plasmid and vaccine production. BsaI is a Type IIS enzyme: it recognises GGTCTC and cuts one base downstream, outside the recognition site, so no scar is left in the product. That makes it the workhorse of Golden Gate assembly and the standard choice for linearising plasmid templates ahead of in vitro transcription.\u003c\/p\u003e\n\u003cp\u003eManufacturing is fully traceable through the process and raw materials. No antibiotics, animal-derived materials or animal-derived excipients are used anywhere in the process. Process-related impurities are controlled and released against specification: host cell protein, residual host DNA, non-specific endonuclease, DNase and RNase activity, microbial limits, bacterial endotoxin, mycoplasma and heavy metals.\u003c\/p\u003e\n\u003cp\u003eThe enzyme digests target DNA in 15 minutes to 1 hour at 37 °C and is heat-inactivated at 80 °C for 20 minutes. It is supplied at 20 U\/µL with six vials of the matching 10× Cut Buffer F, GMP Grade, so no buffer has to be ordered separately. A Certificate of Analysis is issued for every lot.\u003c\/p\u003e\n\u003cp\u003eFor Research Use Only unless qualified by the customer for its intended manufacturing use. Not for use in diagnostic or therapeutic procedures.\u003c\/p\u003e\n\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eSpecifications\u003c\/h2\u003e\n\u003ctable border=\"1\" style=\"border-collapse: collapse; width: 90%;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eParameter\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eSpecification\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eProduct Name\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eBiofargo BsaI, GMP Grade\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eCatalog Number\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eBF-E4001\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eGrade\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eGMP Grade\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eEnzyme Type\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eType IIS restriction endonuclease\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eRecognition \/ Cut Site\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e5′…G G T C T C(N)\u003csub\u003e1\u003c\/sub\u003e…3′\u003cbr\u003e3′…C C A G A G(N)\u003csub\u003e5\u003c\/sub\u003e…5′\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eIsoschizomers\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eEco31I, Bso31I, BspTNI (methylation sensitivities may differ)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eConcentration\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e20 U\/µL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003ePack Size\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e20,000 U (1 mL)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eSupplied Buffer\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e10× Cut Buffer F, GMP Grade, 6 × 1 mL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eReaction Temperature\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e37 °C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eDigestion Time\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e15 minutes to 1 hour\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eHeat Inactivation\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e80 °C for 20 minutes\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eUnit Definition\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eCompletely digest 1 µg of pPIC9K (Dcm−) in 1 hour at 37 °C in a 50 µL reaction\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eProtein Purity\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e≥95% by SDS-PAGE (Coomassie Blue)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eHost Cell Protein\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u0026lt;50 ppm (ChP 2025 Vol. IV, General Chapter 3412)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eResidual Host DNA\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u0026lt;10 copies\/20 U (ChP 2025 Vol. IV, General Chapter 3407, qPCR)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eBacterial Endotoxin\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u0026lt;10 EU\/mg (ChP 2025 Vol. IV, General Chapter 1143, gel-clot)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eMicrobial Limit\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eTotal aerobic count \u0026lt;5 cfu\/mL; yeasts and moulds \u0026lt;5 cfu\/mL (ChP 2025 Vol. IV, General Chapter 1105)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eMycoplasma\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eNegative (LAMP)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eHeavy Metals\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u0026lt;10 ppm (ChP 2025 Vol. IV, General Chapter 0821)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eAnimal-Derived Components\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eNone\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eAntibiotics\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eNone used in manufacture\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eStorage\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e−20 ± 5 °C, valid 24 months\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eShipping\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e≤0 °C (blue ice)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eDocumentation\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003eLot-specific Certificate of Analysis\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eFeatures\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eGMP manufacture with full traceability\u003c\/strong\u003e of process and raw materials, for use as a raw material in mRNA, plasmid and vaccine production.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eType IIS, scarless\u003c\/strong\u003e — cleavage occurs outside the recognition sequence, so no residual bases are left at the cut and the 3′ end of a linearised template is defined by the construct rather than by the enzyme.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eNo animal-derived components and no antibiotics\u003c\/strong\u003e anywhere in the process, which removes two of the questions a raw-material compliance review always asks.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eImpurities released against specification\u003c\/strong\u003e — host cell protein, residual host DNA, non-specific endonuclease, DNase, RNase, microbial limits, endotoxin, mycoplasma and heavy metals, each by a stated pharmacopoeial method.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eBuffer included\u003c\/strong\u003e — six 1 mL vials of 10× Cut Buffer F, GMP Grade ship with the enzyme, so the digest does not depend on a separate buffer order.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eFast, complete digestion\u003c\/strong\u003e in 15 minutes to 1 hour, with heat inactivation at 80 °C for 20 minutes.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eLot-specific Certificate of Analysis\u003c\/strong\u003e with every shipment.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003e24-month shelf life\u003c\/strong\u003e at −20 ± 5 °C.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eApplications\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003ePlasmid linearisation for IVT:\u003c\/strong\u003e Linearise the DNA template before in vitro transcription, where template integrity and a defined 3′ end set the transcript length and translation efficiency.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eGolden Gate and modular assembly:\u003c\/strong\u003e Cut and ligate in one reaction with no scar between parts.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003emRNA manufacture:\u003c\/strong\u003e Supply the upstream digestion step as a GMP-grade raw material with documented impurity control.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003ePlasmid DNA production:\u003c\/strong\u003e Linearise and verify plasmid constructs under a controlled process.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eVaccine and cell therapy manufacture:\u003c\/strong\u003e Use as a raw material where animal-component-free and antibiotic-free sourcing has to be demonstrated.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003e\u003cstrong\u003eProcess development:\u003c\/strong\u003e Run the same enzyme at research scale and at manufacturing scale without changing supplier or revalidating.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eKit Components\u003c\/h2\u003e\n\u003ctable border=\"1\" style=\"border-collapse: collapse; width: 90%;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eComponent\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eVolume\u003c\/strong\u003e\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e\u003cstrong\u003eStorage\u003c\/strong\u003e\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003eBsaI, GMP Grade (20 U\/µL)\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e20,000 U (1 mL)\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e−20 ± 5 °C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"text-align: center;\"\u003e10× Cut Buffer F, GMP Grade\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e6 × 1 mL\u003c\/td\u003e\n\u003ctd style=\"text-align: center;\"\u003e−20 ± 5 °C\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eStorage \u0026amp; Shipping\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\u003cp\u003eShip at ≤0 °C on blue ice.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eStore at −20 ± 5 °C. Valid for 24 months.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eAvoid repeated freeze-thaw cycles; keep the enzyme on ice while setting up reactions.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eA lot-specific Certificate of Analysis is supplied with every shipment.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eProtocol Overview\u003c\/h2\u003e\n\u003col\u003e\n\u003cli\u003e\u003cp\u003eCombine on ice, in order: nuclease-free water to 50 µL, 5 µL of 10× Cut Buffer F, GMP Grade, 1 µg of DNA, and 1 µL of BsaI, GMP Grade (20 U\/µL).\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eMix gently and spin down.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eIncubate at 37 °C for 15 minutes to 1 hour.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eInactivate by heating at 80 °C for 20 minutes.\u003c\/p\u003e\u003c\/li\u003e\n\u003cli\u003e\u003cp\u003eKeep the enzyme below 10% of the total reaction volume to avoid star activity from carried-over glycerol.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ol\u003e\n\u003c\/div\u003e\n\u003cstyle\u003e\n  .bf-rich .josh-faq { background: #fff; color: #000; font-family: \"IBM Plex Sans\", sans-serif; }\n  .bf-rich .josh-faq__inner { padding: 24px 0 40px; max-width: 100%; box-sizing: border-box; }\n  .bf-rich .josh-faq__item { border-bottom: 1px solid #e5e7eb; padding: 0; }\n  .bf-rich .josh-faq__item:first-child { border-top: 1px solid #e5e7eb; }\n  .bf-rich .josh-faq__question { width: 100%; margin: 0; padding: 20px 0; display: flex; align-items: center; justify-content: space-between; gap: 16px; color: #505052; background: none; border: 0; font: 700 16px\/1.4 \"IBM Plex Sans\", sans-serif; text-align: left; cursor: pointer; list-style: none; }\n  .bf-rich .josh-faq__question::-webkit-details-marker { display: none; }\n  .bf-rich .josh-faq__question:hover { color: #505052; }\n  .bf-rich .josh-faq__icon { position: relative; width: 20px; height: 20px; flex: 0 0 20px; transition: transform .25s ease; }\n  .bf-rich .josh-faq__icon::before, .bf-rich .josh-faq__icon::after { position: absolute; content: \"\"; background: currentColor; border-radius: 2px; transition: opacity .25s ease, transform .25s ease; }\n  .bf-rich .josh-faq__icon::before { top: 50%; left: 0; width: 100%; height: 2px; transform: translateY(-50%); }\n  .bf-rich .josh-faq__icon::after { top: 0; left: 50%; width: 2px; height: 100%; transform: translateX(-50%); }\n  .bf-rich .josh-faq__item[open] .josh-faq__icon::after { opacity: 0; transform: translateX(-50%) rotate(90deg); }\n  .bf-rich .josh-faq__answer { max-width: 860px; padding: 0 0 20px; color: #505052; font: 400 16px\/1.6 \"IBM Plex Sans\", sans-serif; }\n  .bf-rich .josh-faq__answer p { margin: 0 0 15px; color: #505052; font: inherit; }\n  .bf-rich .josh-faq__answer p:last-child { margin-bottom: 0; }\n  .bf-rich .josh-faq__answer ul { margin: 8px 0 12px; padding-left: 22px; }\n  .bf-rich .josh-faq__answer li { margin-bottom: 6px; font: inherit; list-style-type: disc; }\n  @media (max-width: 767px) { .bf-rich .josh-faq__inner { padding: 24px 0 40px; } .bf-rich .josh-faq__question { padding: 16px 0; font-size: 15px; } .bf-rich .josh-faq__answer { font-size: 15px; } }\n\u003c\/style\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eFrequently Asked Questions (FAQs)\u003c\/h2\u003e\n\u003cp\u003eCommon questions about Biofargo BsaI, GMP Grade.\u003c\/p\u003e\n\u003cdiv class=\"josh-faq\"\u003e\u003csection class=\"josh-faq__inner\"\u003e\u003cdiv class=\"josh-faq__list\"\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eWhat does GMP Grade mean for this enzyme?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eIt is manufactured and released under GMP specifications, with full traceability of the process and of the raw materials, and with process-related impurities controlled to a written specification. Host cell protein, residual host DNA, non-specific endonuclease, DNase and RNase activity, microbial limits, bacterial endotoxin, mycoplasma and heavy metals are each tested by a stated pharmacopoeial method and reported on the lot Certificate of Analysis.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eAre animal-derived materials or antibiotics used?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eNo. Neither animal-derived materials or excipients nor antibiotics are used anywhere in the manufacturing process. This is normally the first question in a raw-material compliance review, so it is stated on the Certificate of Analysis.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eWhat impurity levels are specified?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eProtein purity is ≥95% by SDS-PAGE. Host cell protein is \u0026lt;50 ppm, residual host DNA \u0026lt;10 copies\/20 U, bacterial endotoxin \u0026lt;10 EU\/mg, heavy metals \u0026lt;10 ppm, total aerobic microbial count \u0026lt;5 cfu\/mL with yeasts and moulds \u0026lt;5 cfu\/mL, and mycoplasma negative by LAMP.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eHow is a digest set up?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eOn ice, combine 5 µL of 10× Cut Buffer F, GMP Grade, 1 µg of DNA, 1 µL of enzyme and nuclease-free water to 50 µL. Mix gently, spin down, incubate at 37 °C for 15 minutes to 1 hour, then inactivate at 80 °C for 20 minutes.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eWhy should the enzyme stay below 10% of the reaction volume?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eThe enzyme storage buffer contains glycerol. Above about 10% of the total reaction volume, the carried-over glycerol can cause star activity. The same logic applies in reverse to contaminants in the DNA: the smaller the reaction, the more strongly salt, EDTA or ethanol in the substrate inhibits digestion.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eWhat can interfere with the digest?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eDNA substrates should carry no phenol, chloroform, ethanol, EDTA, detergent or high salt. All of these reduce enzyme activity, and their effect is proportionally worse in a small reaction volume.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eIs cleavage affected by methylation?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003ePartly. Dam methylation: no effect; Dcm methylation: impaired; CpG methylation: impaired; EcoKI methylation: no effect; EcoBI methylation: no effect. The table on this page gives the full picture; check it against the host the plasmid was grown in.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eDoes it come with buffer?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eYes. Six 1 mL vials of 10× Cut Buffer F, GMP Grade ship with every unit, so the digest does not depend on ordering a buffer separately.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eHow is it stored and shipped?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eIt ships at ≤0 °C on blue ice and is stored at −20 ± 5 °C, where it is valid for 24 months.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eCan you support a DMF filing or a custom GMP enzyme?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eYes. Drug Master File support and custom GMP enzyme manufacture can be arranged. Contact us at contact@biofargo.com with the enzyme, the scale and the regulatory route you need.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eDo you provide a Certificate of Analysis?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eYes. A lot-specific Certificate of Analysis is issued with every shipment and can be sent in advance for a raw-material qualification review. Request it at contact@biofargo.com.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003cdetails class=\"josh-faq__item\"\u003e\u003csummary class=\"josh-faq__question\"\u003eCan it be used for clinical or diagnostic purposes?\u003cspan class=\"josh-faq__icon\" aria-hidden=\"true\"\u003e\u003c\/span\u003e\u003c\/summary\u003e\u003cdiv class=\"josh-faq__answer\"\u003e\u003cp\u003eNo. This product is for research use only and is not intended for human, diagnostic or therapeutic use.\u003c\/p\u003e\u003c\/div\u003e\u003c\/details\u003e\n\u003c\/div\u003e\u003c\/section\u003e\u003c\/div\u003e\n\u003c\/div\u003e\n\u003cscript type=\"application\/ld+json\"\u003e{\"@context\": \"https:\/\/schema.org\", \"@type\": \"FAQPage\", \"mainEntity\": [{\"@type\": \"Question\", \"name\": \"What does GMP Grade mean for this enzyme?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"It is manufactured and released under GMP specifications, with full traceability of the process and of the raw materials, and with process-related impurities controlled to a written specification. Host cell protein, residual host DNA, non-specific endonuclease, DNase and RNase activity, microbial limits, bacterial endotoxin, mycoplasma and heavy metals are each tested by a stated pharmacopoeial method and reported on the lot Certificate of Analysis.\"}}, {\"@type\": \"Question\", \"name\": \"Are animal-derived materials or antibiotics used?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"No. Neither animal-derived materials or excipients nor antibiotics are used anywhere in the manufacturing process. This is normally the first question in a raw-material compliance review, so it is stated on the Certificate of Analysis.\"}}, {\"@type\": \"Question\", \"name\": \"What impurity levels are specified?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Protein purity is \u0026ge;95% by SDS-PAGE. Host cell protein is \u0026lt;50 ppm, residual host DNA \u0026lt;10 copies\/20 U, bacterial endotoxin \u0026lt;10 EU\/mg, heavy metals \u0026lt;10 ppm, total aerobic microbial count \u0026lt;5 cfu\/mL with yeasts and moulds \u0026lt;5 cfu\/mL, and mycoplasma negative by LAMP.\"}}, {\"@type\": \"Question\", \"name\": \"How is a digest set up?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"On ice, combine 5 \u0026#181;L of 10\u0026times; Cut Buffer F, GMP Grade, 1 \u0026#181;g of DNA, 1 \u0026#181;L of enzyme and nuclease-free water to 50 \u0026#181;L. Mix gently, spin down, incubate at 37 \u0026#176;C for 15 minutes to 1 hour, then inactivate at 80 \u0026#176;C for 20 minutes.\"}}, {\"@type\": \"Question\", \"name\": \"Why should the enzyme stay below 10% of the reaction volume?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"The enzyme storage buffer contains glycerol. Above about 10% of the total reaction volume, the carried-over glycerol can cause star activity. The same logic applies in reverse to contaminants in the DNA: the smaller the reaction, the more strongly salt, EDTA or ethanol in the substrate inhibits digestion.\"}}, {\"@type\": \"Question\", \"name\": \"What can interfere with the digest?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"DNA substrates should carry no phenol, chloroform, ethanol, EDTA, detergent or high salt. All of these reduce enzyme activity, and their effect is proportionally worse in a small reaction volume.\"}}, {\"@type\": \"Question\", \"name\": \"Is cleavage affected by methylation?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Partly. Dam methylation: no effect; Dcm methylation: impaired; CpG methylation: impaired; EcoKI methylation: no effect; EcoBI methylation: no effect. The table on this page gives the full picture; check it against the host the plasmid was grown in.\"}}, {\"@type\": \"Question\", \"name\": \"Does it come with buffer?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Yes. Six 1 mL vials of 10\u0026times; Cut Buffer F, GMP Grade ship with every unit, so the digest does not depend on ordering a buffer separately.\"}}, {\"@type\": \"Question\", \"name\": \"How is it stored and shipped?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"It ships at \u0026le;0 \u0026#176;C on blue ice and is stored at \u0026minus;20 \u0026plusmn; 5 \u0026#176;C, where it is valid for 24 months.\"}}, {\"@type\": \"Question\", \"name\": \"Can you support a DMF filing or a custom GMP enzyme?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Yes. Drug Master File support and custom GMP enzyme manufacture can be arranged. Contact us at contact@biofargo.com with the enzyme, the scale and the regulatory route you need.\"}}, {\"@type\": \"Question\", \"name\": \"Do you provide a Certificate of Analysis?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"Yes. A lot-specific Certificate of Analysis is issued with every shipment and can be sent in advance for a raw-material qualification review. Request it at contact@biofargo.com.\"}}, {\"@type\": \"Question\", \"name\": \"Can it be used for clinical or diagnostic purposes?\", \"acceptedAnswer\": {\"@type\": \"Answer\", \"text\": \"No. This product is for research use only and is not intended for human, diagnostic or therapeutic use.\"}}]}\u003c\/script\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eMethylation Effects on Digestion\u003c\/h2\u003e\n\u003ctable style=\"width:100%;border-collapse:collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eDam\u003c\/td\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eDcm\u003c\/td\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eCpG\u003c\/td\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eEcoKI\u003c\/td\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eEcoBI\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eNo effect\u003c\/td\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eImpaired\u003c\/td\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eImpaired\u003c\/td\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eNo effect\u003c\/td\u003e\n\u003ctd style=\"padding:6px 10px;text-align:center;\"\u003eNo effect\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003ch2\u003eNumber of Recognition Sites in Common DNA\u003c\/h2\u003e\n\u003ctable style=\"width:100%;border-collapse:collapse;\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003eλDNA\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003eΦX174\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003epBR322\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003epUC57\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003epUC18\/19\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003eSV40\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003eM13mp18\/19\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003eAdeno2\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003e2\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003e1\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003e0\u003c\/td\u003e\n\u003ctd style=\"padding:6px 8px;text-align:center;\"\u003e18\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/tbody\u003e\n\u003c\/table\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eRelated Products\u003c\/h2\u003e\n\u003cp\u003eThe Biofargo GMP-grade restriction endonuclease range covers both scarless Type IIS sites and the conventional backbone sites, so a change of plasmid backbone does not mean a change of supplier or a second raw-material qualification.\u003c\/p\u003e\n\u003cul\u003e\n  \u003cli\u003e\u003cp\u003e\u003ca href=\"\/products\/biofargo-xbai-gmp-grade\"\u003e\u003cstrong\u003eBiofargo XbaI, GMP Grade\u003c\/strong\u003e\u003c\/a\u003e — Type II, palindromic site; 20 U\/µL, 20,000 U.\u003c\/p\u003e\u003c\/li\u003e\n  \u003cli\u003e\u003cp\u003e\u003ca href=\"\/products\/biofargo-bspqi-gmp-grade\"\u003e\u003cstrong\u003eBiofargo BspQI, GMP Grade\u003c\/strong\u003e\u003c\/a\u003e — Type IIS, scarless cleavage; 10 U\/µL, 10,000 U.\u003c\/p\u003e\u003c\/li\u003e\n  \u003cli\u003e\u003cp\u003e\u003ca href=\"\/products\/biofargo-xhoi-gmp-grade\"\u003e\u003cstrong\u003eBiofargo XhoI, GMP Grade\u003c\/strong\u003e\u003c\/a\u003e — Type II, palindromic site; 20 U\/µL, 20,000 U.\u003c\/p\u003e\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c\/div\u003e\n\u003cdiv class=\"bf-rich\"\u003e\n\u003ch2\u003eDocuments\u003c\/h2\u003e\n\u003cp\u003e\u003ca target=\"_blank\" rel=\"noopener\" href=\"https:\/\/biofargo.com\/pages\/contact\"\u003e\u003cstrong\u003eRequest Datasheet\u003c\/strong\u003e\u003c\/a\u003e\u003c\/p\u003e\n\u003c\/div\u003e\n","brand":"Biofargo","offers":[{"title":"Default Title","offer_id":53432385175733,"sku":"BF-E4001","price":1050.0,"currency_code":"USD","in_stock":true}],"url":"https:\/\/biofargo.com\/products\/biofargo-bsai-gmp-grade","provider":"Biofargo","version":"1.0","type":"link"}