User Manual
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1. lt gt MCLAB Molecular Cloning Laboratories User Manual Version 1 0 Revision Date 08 30 2013 Product name 17 Exonuclease Cat T7G6E 1 00 T7G6E 200 T7G6E 300 T7G6E OEM Description T7 Exonuclease is similar to Lambda Exonuclease in that it catalyzes the stepwise hydrolysis of duplex DNA from the 5 termini liberating 5 mononucleotides However unlike Lambda Exonuclease the enzyme has low processivity and it will remove both 5 hydroxyl and 5 phosphoryl termini T7 Exonuclease hydrolyzes duplex DNA non processively in the 5 gt 3 direction from both 5 phosphoryl or 5 hydroxyl nucleotides by liberating oligonucleotides as well as mononucleotides until about 50 of the DNA is acid soluble Source Purified from an E coli strain containing a TYB12 intein fusion Storage Conditions 10 mM Tris HCI 5 mM DTT 0 1 mM EDTA 50 Glycerol pH 8 0 25 C Application Controlled stepwise digestion of double stranded DNA from the 5 termini Generating ssDNA templates for sequencing via the chain termination method Supplied with 10X T7 Exonuclease Buffer 10X T7 Exonuclease Buffer 500 mM Potassium Acetate 200 mM Tris acetate 100 mM Magnesium Acetate 10 mM DTT pH 7 9 25 C Unit Definition One unit is the amount of enzyme required to release 1 nmol of acid soluble nucleotide in 15 min at 37 C under standard assay conditions Recommended Storage Conditions 20 C 1 650 872. 2 0245 www mclab com 1 Reference 1 Kerr C and Sadowski P D 1972 J Biol Chem 247 311 318 2 Thomas K R and Olivera B M 1978 J Biol Chem 253 424 429 3 Ausubel F M Brent R Kingston R E Moore D D Seidman J G Smith J A and Struhl K 1987 Cur rent Protocols in Molecular Biology John Wiley and Sons Inc 4 Shon M Germino J and Bastia D 1982 J Biol Chem 257 13823 13827 5 Nikiforov T T Rendle R B Goelet P Rogers Y H Kotewicz 6 M L Anderson S Trainor G L and Knapp M R 1994 Nucl Acids Res 22 20 4167 4175 7 Kornberg A and Baker T 1991 DNA Replication Second Edition 591 2 lt MCLAB
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