0 item(s)
View cart
You have no items in your shopping cart.
$1,050.00
Sku: BF-E4001
Availability:
AVAILABLE
In stock & estimated to ship in 3-7 days by October 13, 2026
Description
Biofargo BsaI, GMP Grade is a restriction endonuclease produced by recombinant expression in E. coli 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.
Manufacturing 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.
The 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.
For Research Use Only unless qualified by the customer for its intended manufacturing use. Not for use in diagnostic or therapeutic procedures.
Specifications
| Parameter | Specification |
| Product Name | Biofargo BsaI, GMP Grade |
| Catalog Number | BF-E4001 |
| Grade | GMP Grade |
| Enzyme Type | Type IIS restriction endonuclease |
| Recognition / Cut Site | 5′…G G T C T C(N)1…3′ 3′…C C A G A G(N)5…5′ |
| Isoschizomers | Eco31I, Bso31I, BspTNI (methylation sensitivities may differ) |
| Concentration | 20 U/µL |
| Pack Size | 20,000 U (1 mL) |
| Supplied Buffer | 10× Cut Buffer F, GMP Grade, 6 × 1 mL |
| Reaction Temperature | 37 °C |
| Digestion Time | 15 minutes to 1 hour |
| Heat Inactivation | 80 °C for 20 minutes |
| Unit Definition | Completely digest 1 µg of pPIC9K (Dcm−) in 1 hour at 37 °C in a 50 µL reaction |
| Protein Purity | ≥95% by SDS-PAGE (Coomassie Blue) |
| Host Cell Protein | <50 ppm (ChP 2025 Vol. IV, General Chapter 3412) |
| Residual Host DNA | <10 copies/20 U (ChP 2025 Vol. IV, General Chapter 3407, qPCR) |
| Bacterial Endotoxin | <10 EU/mg (ChP 2025 Vol. IV, General Chapter 1143, gel-clot) |
| Microbial Limit | Total aerobic count <5 cfu/mL; yeasts and moulds <5 cfu/mL (ChP 2025 Vol. IV, General Chapter 1105) |
| Mycoplasma | Negative (LAMP) |
| Heavy Metals | <10 ppm (ChP 2025 Vol. IV, General Chapter 0821) |
| Animal-Derived Components | None |
| Antibiotics | None used in manufacture |
| Storage | −20 ± 5 °C, valid 24 months |
| Shipping | ≤0 °C (blue ice) |
| Documentation | Lot-specific Certificate of Analysis |
Features
GMP manufacture with full traceability of process and raw materials, for use as a raw material in mRNA, plasmid and vaccine production.
Type IIS, scarless — 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.
No animal-derived components and no antibiotics anywhere in the process, which removes two of the questions a raw-material compliance review always asks.
Impurities released against specification — host cell protein, residual host DNA, non-specific endonuclease, DNase, RNase, microbial limits, endotoxin, mycoplasma and heavy metals, each by a stated pharmacopoeial method.
Buffer included — 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.
Fast, complete digestion in 15 minutes to 1 hour, with heat inactivation at 80 °C for 20 minutes.
Lot-specific Certificate of Analysis with every shipment.
24-month shelf life at −20 ± 5 °C.
Applications
Plasmid linearisation for IVT: Linearise the DNA template before in vitro transcription, where template integrity and a defined 3′ end set the transcript length and translation efficiency.
Golden Gate and modular assembly: Cut and ligate in one reaction with no scar between parts.
mRNA manufacture: Supply the upstream digestion step as a GMP-grade raw material with documented impurity control.
Plasmid DNA production: Linearise and verify plasmid constructs under a controlled process.
Vaccine and cell therapy manufacture: Use as a raw material where animal-component-free and antibiotic-free sourcing has to be demonstrated.
Process development: Run the same enzyme at research scale and at manufacturing scale without changing supplier or revalidating.
Kit Components
| Component | Volume | Storage |
| BsaI, GMP Grade (20 U/µL) | 20,000 U (1 mL) | −20 ± 5 °C |
| 10× Cut Buffer F, GMP Grade | 6 × 1 mL | −20 ± 5 °C |
Storage & Shipping
Ship at ≤0 °C on blue ice.
Store at −20 ± 5 °C. Valid for 24 months.
Avoid repeated freeze-thaw cycles; keep the enzyme on ice while setting up reactions.
A lot-specific Certificate of Analysis is supplied with every shipment.
Protocol Overview
Combine 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).
Mix gently and spin down.
Incubate at 37 °C for 15 minutes to 1 hour.
Inactivate by heating at 80 °C for 20 minutes.
Keep the enzyme below 10% of the total reaction volume to avoid star activity from carried-over glycerol.
Frequently Asked Questions (FAQs)
Common questions about Biofargo BsaI, GMP Grade.
What does GMP Grade mean for this enzyme?
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.
Are animal-derived materials or antibiotics used?
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.
What impurity levels are specified?
Protein purity is ≥95% by SDS-PAGE. Host cell protein is <50 ppm, residual host DNA <10 copies/20 U, bacterial endotoxin <10 EU/mg, heavy metals <10 ppm, total aerobic microbial count <5 cfu/mL with yeasts and moulds <5 cfu/mL, and mycoplasma negative by LAMP.
How is a digest set up?
On 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.
Why should the enzyme stay below 10% of the reaction volume?
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.
What can interfere with the digest?
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.
Is cleavage affected by methylation?
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.
Does it come with buffer?
Yes. 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.
How is it stored and shipped?
It ships at ≤0 °C on blue ice and is stored at −20 ± 5 °C, where it is valid for 24 months.
Can you support a DMF filing or a custom GMP enzyme?
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.
Do you provide a Certificate of Analysis?
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.
Can it be used for clinical or diagnostic purposes?
No. This product is for research use only and is not intended for human, diagnostic or therapeutic use.
Methylation Effects on Digestion
| Dam | Dcm | CpG | EcoKI | EcoBI |
| No effect | Impaired | Impaired | No effect | No effect |
Number of Recognition Sites in Common DNA
| λDNA | ΦX174 | pBR322 | pUC57 | pUC18/19 | SV40 | M13mp18/19 | Adeno2 |
| 2 | 0 | 1 | 1 | 1 | 0 | 0 | 18 |
Related Products
The 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.
Biofargo XbaI, GMP Grade — Type II, palindromic site; 20 U/µL, 20,000 U.
Biofargo BspQI, GMP Grade — Type IIS, scarless cleavage; 10 U/µL, 10,000 U.
Biofargo XhoI, GMP Grade — Type II, palindromic site; 20 U/µL, 20,000 U.
Documents
When can I expect my order to ship?
Most orders are filled and shipped within 2-3 business days from the time they are received.
Our standard shipping usually take 2-5 days.
We also provide express shippping for time-sensitive deliveries.
Email contact@biofargo.com if you have any requirements.
