B3100
BsuRI (HaeIII)
Size
£384.00
- SKU:
- B3100
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Extra Details:
- 5'-G G^C C-3' 3'-C C^G G-5' Source: Bacillus subtilis R Diluent Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Thermal Inactivation: BsuRI is inactivated by incubation at 80°C for 20 minutes. Storage Buffer: 10mM Tris-HCl, pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Supplied with: R1625: Restriction Enzyme Buffer A, 10X: Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. R1625-04: Dilute to 1X for 100% BsuRI digestion. 10mM Tris-HCl, pH 8.5, 10mM MgCl2, 100mM KCl and 0.1mg/ml BSA. Incubation Temperature: 37°C Unit Definition: One unit is defined as the amount of BsuRI required to digest 1ug of lambda DNA in 1 hour at 37°C in 50ul of assay buffer. Methylation Effects: Dam, EcoKl, EcoBl: never overlaps - no effect. Dcm, CpG: may overlap - no effect. Protocol for Digestion: Add: Nuclease free water: 16ul R1625-04: 2ul DNA (0.5-1ug/ml): 1ul B3100: 0.5-2ul Mix gently and spin down for a few seconds. Incubate at 37ºC for 1-16 hours. Protocol for Digestion Directly after Amplification: Add: PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA) Nuclease free water: 18ul R1625-04: 2ul B3100: 1-2ul Mix gently and spin down for a few seconds. Incubate at 37ºC for 1-16 hours. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BsuRI. Ligation/Recleavage (L/R) Assay: The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of BsuRI for 4 hours. Stability during Prolonged Incubation: A minimum of 0.1units of BsuRI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. Number of Recognition Sites in DNA: Lambda: 149 PhiX174, pUC18/19: 11 M13mp18/19: 15 pBR322: 22 pUC57: 13 pTZ19R/U: 12 Storage and Stability: Aliquot and store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
- Shipping Conditions:
- Blue Ice