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T4427

Sigma-Aldrich

Terminal Transferase from calf thymus

buffered aqueous glycerol solution

Synonym(s):

TdT, Terminal Deoxynucleotidyl Transferase

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About This Item

CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
UNSPSC Code:
12352204

grade

Molecular Biology
for molecular biology

form

buffered aqueous glycerol solution

mol wt

60 kDa

concentration

>5000 U/mL

UniProt accession no.

foreign activity

Exonuclease and endonuclease, free

storage temp.

−20°C

Gene Information

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General 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.

Application

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

Components

Terminal transferase is supplied as a solution of 50 mM potassium phosphate (pH 7.4), 1 mM 2-mercaptoethanol and 50% glycerol (v/v).

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.

Analysis Note

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.

Pictograms

Health hazard

Signal Word

Danger

Hazard Statements

Precautionary Statements

Hazard Classifications

Resp. Sens. 1

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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C P Tu et al.
Gene, 10(2), 177-183 (1980-07-01)
Cordycepin-5'-triphosphate (3'-deoxyadenosine-5'-triphosphate) can be incorporated into the 3'-ends of DNA fragments using terminal deoxynucleotidyl transferase from calf thymus (Bollum, 1974). Because cordycepin-5'-monophosphate lacks a 3'-OH group, only a single residue is incorporated. Furthermore, DNA molecules that contain cordycepin-5'-monophosphate at their
Marcel Hollenstein et al.
Bioorganic & medicinal chemistry letters, 22(13), 4428-4430 (2012-05-29)
Pyrene-deoxynucleoside triphosphates (dPTPs), varying by the positioning of the aromatic system, were synthesized. Their ability to function as substrates for the Klenow fragment of Escherichia coli DNA polymerase I and the TdT polymerase was assessed. The dPTPs are all equally
Yuan Zhang et al.
Cellular immunology, 274(1-2), 19-25 (2012-04-03)
Secondary rearrangements of the T cell receptor (TCR) represent a genetic correction mechanism which changes T cell specificity by re-activating V(D)J recombination in peripheral T cells. Murine T-cell hybridoma A1.1 was employed to investigate whether antigenic stimulation induced re-expression of
Botao Zhao et al.
Acta biochimica et biophysica Sinica, 44(2), 129-135 (2011-12-23)
MicroRNAs (miRNAs) constitute a critically important class of non-translated, small RNAs, which post-transcriptionally regulate gene expression via one of the multiple mechanisms. To profile miRNA expression, microarrays have been extensively applied to the high-throughput detection of miRNAs. Here, we described
Maniatis, T., et al.
Molecular Cloning: A Laboratory Manual, 148-148 (1989)

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