NcoI

Product Overview

• Article Name: NcoI
• Biozol Catalog Number: USB-N0554
• Supplier Catalog Number: N0554
• Alternative Catalog Number: USB-N0554-1000,USB-N0554-2500
• Manufacturer: US Biological
• Category: Molekularbiologie

Product Description

5-C C A T G G-3 3-G G T A C C-5 Source: Nocardia corallina Concentration: 10u/ul Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Supplied With: R1625: Restriction Enzyme Buffer A, 10X Supplied as a liquid. 1x contains 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Storage Buffer: Supplied as a liquid in 10mM Tris-HCl, pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Dilution Buffer: 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Double Digests: Restriction Enzyme Buffer A, 10X, is provided to simplify buffer selection for double digests. Diluted to 1X, the formulation is 33mM Tris-acetate, pH 7.9, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Incubate at 37C. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with NcoI. Ligation Recleavage (L/R) Assay: The ligation and recleavage assay was replaced with LO test after validating experiments showed LO test ability to trace nuclease and phosphatase activities with sensitivity that is higher than L/R by a factor of 100. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded or double-stranded labeled oligonucleotide was observed after incubation with 10 units of NcoI for 4 hours. Blue White Cloning Assay: The B/W assay was replaced with LO test after validating experiments showed LO test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test. Thermal Inactivation: NcoI is inactivated by incubation at 65C for 20min. Digestion of Agarose-embedded DNA: A minimum of 5 units of NcoI is required for digestion of 1ug of agarose-embedded lambda DNA in 16 hours. Methylation Effects on Digestion: Dam, Dcm, CpG, EcoKI, EcoBI: Never overlaps, no effect Compatible Ends: Afl III, PscI, BtgI, Eco130I, FatI, Pag I Number of Recognition Sites in DNA: Lambda: 4, PhiX174, M13mp18/19, pBR322, pUC18/19, pUC57, pTZ19R/U: 0 Stability During Prolonged Incubation: A minimum of 0.1 units of NcoI is required for complete digestion of 1ug of DNA in 16 hours at 37C. Protocol for Digestion: Add Nuclease free water (16ul) R1625 (2ul) DNA (0.5-1ug/ml) (1ul) N0554 (0.5-2ul) Mix gently and spin down for a few seconds. Incubate at 37C for 1-16 hours Protocol for Digestion Directly after Amplification: Add PCR Reaction Mixture (10ul) (~0.1-0.5ug of DNA)) Nuclease free water (18ul) R1625 (2ul) N0554 (1-2ul) Mix gently and spin down for a few seconds. Incubate at 37C for 1-16 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.