Rapid DNA Digestion 5-15Min Enzyme BsaI Biochemical Reagent CE Approved

Enzymes BsaI

enzymes are a series of engineered restriction enzymes for rapid DNA digestion in 5-15 min.  enzymes can be used to digest plasmid, genomic and viral DNA as well as PCR products. All  enzymes show superior activity in the universal buffer, so simplified the enzymatic digestion reaction system. Moreover, the  enzymes provides excellent enzyme redundancy, allowing easy digestion of substrate excess or difficult templates.

Shipping and Storage

The components are shipped with ice pack and can be stored at -20°C for 2 years.

Cutting Site

5'-GGTCTC(N)1↓-3'

3'-CCAGAG(N)5↑-5'

Recommended Reaction Conditions

1× FuniCut™ Buffer; Incubate at 37℃.

Thermal Inactivation

Incubation at 80℃ for 20 min.

Quality Control

1. Activity definition: 1 μg pPIC9K DNA was completely digested with 1 μL of the enzyme in 15 min at 37℃ in a total reaction volume of 20 μL.

2. Prolonged Incubation/Star Activity Assay: No detectable degradation of 1 μg pPIC9K DNA due to nuclease contamination or star activity occurred during incubation with 1 μL of FuniCut™ BsaI for 3 hour at 37℃. Longer incubation may result in star activity.

3. Ligation and Recleavage (L/R) Assay: After digested with 1μL enzyme under optimal condition, the recycle product from digestion can be ligated under T4 DNA Ligase at 22℃. The ligation product can be recleavaged by the enzyme.

4. Nonspecific Endonuclease Activity: Incubation of 1 μL enzyme with 1 μg supercoiled plasmid DNA for 4 hours at 37℃ resulted in no change of the supercoiled condition.

Instructions

1. Protocol for Fast Digestion of different DNA

1.1 Combine the following reaction components on ice in the order indicated:

[Note]: *For purified PCR products. Amount of 10× FuniCut™ Buffer may be reduced to 2 μL due to the remaining ionic strength in the unpurified PCR products. PCR product was recommended to be purified prior digestion When PCR Product will be used for cloning.

1.2 Mix gently (do not vortex) and spin down.

1.3 Incubate at 37℃ for 15 min (plasmid DNA) or for 15-30 min (PCR product) or for 30-60 min (genomic DNA).

1.4 Optional: Inactivate the enzyme by heating for 20 min at 80℃.

2. Double and Multiple Digestion of DNA

2.1 Use 1 μL of each enzyme and scale up the reaction conditions appropriately.

2.2 The combined volume of the enzymes in the reaction mixture should not exceed 1/10 of the total reaction volume.

2.3 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.

3. Scaling up Plasmid DNA Digestion Reaction

[Note]: Incubation in a water thermostat, metal thermostat or sand thermostat. Increase the incubation time if the total reaction volume exceeds 20 μL.

Number of Recognition Sites in DNA