|R0527L||5,000 units||5,000 units/ml|
|R0527S||1,000 units||5,000 units/ml|
Bacillus stearothermophilus (C. Polisson)
Activity in NEBuffers:
NEBuffer 1: 0%
NEBuffer 2: 50%
NEBuffer 3: 100%
NEBuffer 4: 10%
When using a buffer other than the optimal (supplied) NEBuffer, it may be necessary to add more enzyme to achieve complete digestion.
dam methylation: Not sensitive
dcm methylation: Not sensitive
CpG methylation: Not sensitive
More information about: Methylation Sensitivity
Activity at 37°C:
80°C for 20 minutes
Survival in a Reaction:
(+ +) Intermediate activity. Suitable for extended digestion, but < 8 hours.
More information about: Extended Digests with Restriction Enzymes
Reaction & Storage Conditions
1X NEBuffer 3
Incubate at 65°C.
1X NEBuffer 3:
50 mM Tris-HCl
100 mM NaCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.9 @ 25°C
One unit is defined as the amount of enzyme required to digest 1 µg of ΦX174 DNA in 1 hour at 65°C in a total reaction volume of 50 µl.
Unit Assay Substrate:
10 mM Tris-HCl
400 mM NaCl
1 mM Dithiothreitol
0.1 mM EDTA
200 µg/ml BSA
pH 7.4 @ 25°C
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Ligation and Re-cutting:
After a 10-fold overdigestion with BsrI, > 95% of the DNA fragments can be ligated with T4 DNA Ligase (at a 5' termini concentration of 1-2 μM) at 16ºC. Of these ligated fragments, > 95% can be recut with BsrI.
A 50 μl reaction containing 1 μg of ΦX174 DNA and 50 units of BsrI incubated for 16 hours at 65ºC resulted in a DNA pattern free of detectable nuclease degradation as determined by agarose gel electrophoresis.
Incubation of a 50 μl reaction containing 100 units of BsrI with 1 μg of a mixture of single and double-stranded [3H] E. coli DNA (205 cpm/μg) for 4 hours at 65ºC released < 0.2% of the total radioactivity.
Reagents Sold Separately
Q1: Are there any known sequence errors at the recognition site in a database?
A1: Yes, BsrI cleaves Bluescript 13 times not once as previously reported in Stratagene catalog.
Q2: Does BsrI have trouble cleaving PCR products?
A2: Yes. PCR products should be phenol extracted or column purifed, and EtOH precipitated, before cleaving with BsrI.
Q3: What is the activity of BsrI at 37°C?
A3: Incubation at 37°C results in 20% activity.