Acc65I (KpnI*)

Product Overview

• Article Name: Acc65I (KpnI*)
• Biozol Catalog Number: USB-A0200
• Supplier Catalog Number: A0200
• Alternative Catalog Number: USB-A0200-1000,USB-A0200-5000
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-G G T A C C-3 3-C C A T G G-5 Unlike KpnI, Acc65I produces DNA fragments with a 4-base 5-extension. Concentration: 10 units/ul Source: Acinetobacter aceti 655 Recommended Conditions: Storage Buffer: 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM sodium chloride, 0.1mg/ml BSA. Incubate at 37C. Diluent Buffer: 10mM Tris-HCl, pH 7.4 at 25C, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Storage Buffer: 10mM Tris-HCl, pH 7.4 at 25C, 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Quality Control: Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with Acc65I. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with Acc65I approximately 90% of the DNA fragments can be ligated at a 5-termini concentration of 0.01uM. More than 90% of these can be recut. Labeled Oligonucleotide (LO) Assay (endonuclease, exonuclease and phosphatase): No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours. Blue/White Cloning Assay: pUC57 was digested at a unique site with 10 units of enzyme for 16 hours. After religation and transformation 0.3% of white colonies were detected. Methylation Effects: Acc65I cleaves the hemi-methylated sequence GGTACm5C. Cleavage is blocked when both strands are methylated at positions: 5_ GGTACm5C_3 3_m5CCATG G_5 [Overlapping Dcm or CG methylation may influence DNA cleavage] Stability during Prolonged Incubation: A minimum of 0.3 units of enzyme are required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Thermal Inactivation: Enzyme is inactivated by incubation at 65C for 20min. Compatible Ends: BshNI, Bsp1407I, Pfl23II, TatI Number of Recognition Sites in DNA: Lambda 2 PhiX174 0 M13mp18/19 1 pBR322 0 pUC18/19 1 pUC57 1 pTZ19R/U 1 pBluescriptIIKS(-/+) 1 pBluescriptIISK(-/+) 1 pACYC177 0 pACYC184 0 Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA-BamHI fragments in 1 hour at 37C in 50ul of assay buffer.