BseLI (BslI)

Product Overview

• Article Name: BseLI (BslI)
• Biozol Catalog Number: USB-B2908
• Supplier Catalog Number: B2908
• Alternative Catalog Number: USB-B2908-500
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

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 55C 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 55C. Thermal Inactivation: BseLI is inactivated by incubation at 80C 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 X174: 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 4C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20C. Aliquots are stable for 6 months after receipt at -20C. 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.