Description
Bovine terminal transferase (TdT) is a primer-dependent polymerase that catalyzes the addition of deoxynucleotides to the 3'-OH terminus of DNA molecules with the release of inorganic phosphate. TdT reacts preferentially with either single-stranded DNA molecules or double-stranded-DNA with 3' overhangs, but procedures have been developed to label blunt ends or 3'-recessive ends. In a reaction mixture, the divalent ion (Co2+, Mn2+, Mg2+) will influence purine and pyrimidine polymerization rate. Activities of TdT are also affected by the bases (dATP, dCTP, dGTP and dTTP) present.
Applications
Suitable for: • Addition of homopolymers to vectors, inserts and cDNA for cloning • Labeling the 3′-end of double-and single-stranded DNA with non-radioactive or radioactive labels • Carrying out in vitro mutagenesis by adding single nucleotides to DNA • Use in TUNEL assays
Product Overview
Bovine terminal transferase (TdT) is a primer-dependent polymerase that catalyzes the addition of deoxynucleotides to the 3′-OH terminus of DNA molecules with the release of inorganic phosphate. TdT reacts preferentially with either single-stranded DNA molecules or double-stranded-DNA with 3′ overhangs, but pr ocedures have been developed to label blunt ends or 3′-recessive ends. In a reaction mixture, the divalent ion (Co2+, Mn2+, Mg2+) will influence purine and pyrimidine polymerization rate. Activities of TdT are also affected by the bases (dATP, dCTP, dGTP and dTTP) present.
Form
buffered aqueous glycerol solution
Enzyme Commission Number
EC 2.7.7.31
Molecular Weight
mol wt 60 kDa
Unit Definition
One unit will incorporate 1 nanomole of dATP into acid-precipitable material in one hour at 37°C using d (pT)6 as primer.
Warnings
Activity assay uses 200 mM potassium cacodylate, pH 7.2, 4 mM MgCl2, 1 mM 2-mercaptoethanol, 1 mM 3H-dATP, 70 μM d (pT)6, 37°C.
Function
DNA binding; DNA nucleotidylexotransferase activity; DNA-directed DNA polymerase activity
Synonyms
DNA nucleotidylexotransferase; terminal deoxyribonucleotidyltransferase; terminal addition enzyme; addase; deoxynucleotidyl terminal transferase; deoxyribonucleic acid nucleotidyltransferase; deoxyribonucleic nucleotidyltransferase; terminal deoxynucleotide transferase; TdT; EC 2.7.7.31; 9027-67-2
Download Datasheet