ApaI

Product Overview

• Article Name: ApaI
• Biozol Catalog Number: USB-A2298-63
• Supplier Catalog Number: A2298-63
• Alternative Catalog Number: USB-A2298-63-5000
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-G G G C C C-3 3-C C C G G G-5 Concentration: 10u/ul Source: Acetobacter pasteurianus Buffer: 10mM Tris-HCl (pH 7.5), 10mM MgCl2 and 0.1mg/ml BSA. Incubate at 37C. (Incubation at 30C results in a 2-fold increase in activity.) Diluent Buffer: 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol. For longer periods-the Storage Buffer should be used. Storage Buffer: 10mM Tris-HCl (pH 7.4 at 25C), 50mM NaCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 320-fold overdigestion (20u/ug lambda DNA x 16 hours) with ApaI. Ligation/Recutting Assay After 50-fold overdigestion (3u/ug DNA x 17 hours) with ApaI, more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 0.02uM. 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. Blue/White Cloning Assay pUC57 was digested at a unique site with 10units of enzyme for 16 hours. After religation and transformation 0.3% of white colonies were detected. Methylation Effects: Sequence GGGCCm5C is cleaved approximately 150 times slower than the unmethylated one. Overlapping Dcm or CG methylation may influence DNA cleavage. Stability during Prolonged Incubation: A minimum of 0.2units of enzyme is required for complete digestion of 1ug of DNA in 16 hours at 37C. Thermal Inactivation: Enzyme is inactivated by incubation at 65C for 20min. Digestion of Agarose-embedded DNA: A minimum of 5units of enzyme is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Compatible Ends: GGGCC C-BseSI, GGGCC C-Eco24I, GGGCC C-SduI Number of Recognition Sites in DNA: Lambda: 1 PhiX174: 0 M13mp18/19: 0 pBR322: 0 pUC18/19: 0 pUC57: 1 pTZ19R/U: 0 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-CpoI fragments in 1 hour at 37C in 50ul of assay buffer.