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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

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

signalword

Danger

hcodes

Hazard Classifications

Resp. Sens. 1

Storage Class

11 - Combustible Solids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)


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D G Remick et al.
The American journal of pathology, 136(1), 49-60 (1990-01-01)
Tumor necrosis factor-alpha (TNF) has been implicated strongly as a principal mediator in the pathogenesis of septic shock. The authors investigated the in vivo production of TNF in CBA/J and CD-1 mice that had been primed by an intraperitoneal injection
Edward A Motea et al.
ACS chemical biology, 7(6), 988-998 (2012-03-07)
Acute lymphoblastic leukemia (ALL) is the most common type of childhood cancer, presenting with approximately 5,000 new cases each year in the United States. An interesting enzyme implicated in this disease is terminal deoxynucleotidyl transferase (TdT), a specialized DNA polymerase
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
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

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