RsaI

Product Overview

• Article Name: RsaI
• Biozol Catalog Number: USB-R9300
• Supplier Catalog Number: R9300
• Alternative Catalog Number: USB-R9300-1000,USB-R9300-5000
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-G T A C-3 3-C A T G-5 Source: Rhodopseudomonas sphaeroides Concentration: 10u/ul Form: Supplied as a liquid in 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol Restriction Buffer: (R1625) Supplied as a liquid in 33mM Tris-acetate (pH 7.9), 10mM magnesium acetate, 66mM potassium acetate and 0.1mg/ml BSA. Incubate at 37C Diluent Buffer: Dilute with 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. Thermal Inactivation: Rsal is inactivated by incubation at 80C for 20min. Number of Recognition Sites in DNA Lambda: 113 PhiX174: 11 M13mp18/19: 19 pBR322: 3 pUC18/19: 3 pUC57: 3 pTZ19R/U: 2 Unit Definition: One unit is defined as the amount of RsaI required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Recommended Protocol for Digestion: Add: Nuclease free water: 16ul 10X Restriction Buffer: 2ul DNA (0.5-1ug/ul): 1ul Rsal: 0.5-2ul Mix gently, spin down for a few seconds. Incubate at 37C for 1-16 hours. Protocol for Digestion of PCR products directly after amplification: Add: PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA) Nuclease free water: 18ul 10X Restriction Buffer: 2ul Rsal: 1-2ul Mix gently, spin down for a few seconds. Incubate at 37C for 1-16 hours. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion with RsaI (20u/ug lambda DNA x 16 hours). Ligation Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with RsaI, more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 0.5uM. More than 95% of these can be recut. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded or double-stranded labeled oligonucleotide occured during incubation with 10 units of Rsal for 4 hours. Methylation Effects: Dam: never overlaps-no effect Dcm: never overlaps-no effect CpG: may overlap-cleavage impaired EcoKI: never overlaps-no effect EcoBI: never overlaps-no effect Stability during Prolonged Incubation: A minimum of 0.2 units of RsaI is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C.