B2908
BseLI (BslI)
Size
£307.00
- SKU:
- B2908
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Extra Details:
- 5'-C C N N N N N^N N G G-3' 3'-G G N N^N N N N N C C-5' Source: Bacillus stearothermophilus LK 3–551 Supplied With: R1625: Restriction Enzyme Buffer A, 10X Concentration: 10u/ul Unit Definition: One unit is defined as the amount of BseLI required to digest 1ug of lambda DNA dcm-in 1 hour at 55°C in 50ul of assay buffer. Diluent Buffer: For short term storage (3-4 weeks)- dilute with 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Storage Buffer: For long term storage: 10mM Tris-HCl pH 7.4, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol. R1625: Restriction Enzyme Buffer A, 10X: Suplied as a liquid in 33mM Tris-acetate pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Enzyme Properties: Methylation Effects: Dam: never overlaps - no effect Dcm: may overlap - cleavage impaired CpG: may overlap - cleavage impaired EcoKl: never overlaps - no effect EcoBl: never overlaps - no effect Stability during Prolonged Incubation: A minimum of 0.1units of BseLI is required for complete digestion of 1ug of lambda DNA in 16 hours at 55°C. Thermal Inactivation: BseLI is inactivated by incubation at 80°C for 20min. Inactivation Procedure: To prepare the digested DNA. for electrophoresis: 1. Stop the digestation rection by adding 0.5M EDTA, pH8, to achieve a 20mM final concentration. Mix thoroughly, add an electrophoresis loading dye and load onto gel. To prepare DNA suitable for further enzymatic reactions: 1. Extract with phenol/ chloroform, precipitate with ethanol or isopropanol, wash the pellet with 75% cold ethanol and let air dry. 2. Dissolve DNA in either nuclease free water, TE buffer or a buffer suitable for further applications. Check the DNA concentration in the solution. Number of Recognition Sites in DNA: Lambda: 176 M13mp18/19: 17 pBR322: 20 pUC18/19, pUC57: 6 pTZ19R/U: 7 ϕX174: 19 Note: BseLI cleavage is impaired by overlapping dcm methylation. To avoid dcm methylation, use a dam, dcm strain such as GM2163. Quality Control: Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with BseLI. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with BseLI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 1.6uM. More than 95% of these can be recut. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing.. 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