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Roche

Terminal Transferase

from Calf Thymus, recombinant, E. coli

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8000 units

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$301.00
24000 units
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$723.00
$636.24

About This Item

EC Number:
2.7.7.31 (BRENDA, IUBMB)
NACRES:
NA.21
UNSPSC Code:
12352204
Recombinant:
expressed in E. coli

$301.00

Available to ship TODAYDetails

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recombinant

expressed in E. coli

form

solution

usage

sufficient for 20 reactions (03333566001), sufficient for 60 reactions (03333574001)

packaging

pkg of 24,000 U (03333574001 [400 U per reaction]), pkg of 8,000 U (03333566001 [400 U per reaction])

manufacturer/tradename

Roche

application(s)

genomic analysis

storage temp.

−20°C

General description

Terminal Transferase catalyzes the template independent addition of deoxy- and dideoxynucleoside triphosphates to the 3′-OH ends of double and single-stranded DNA fragments, and oligonucleotides. Terminal Transferase incorporates digoxigenin-, biotin-, and fluorochrome-labeled deoxy- and dideoxynucleoside triphosphates as well as radioactively labeled deoxy- and dideoxynucleoside triphosphates. The supplied 5x-concentrated reaction buffer allows the optimal tailing of all types of double-stranded DNA ends: blunt ended, with 3′ overhang, or with 5′ overhang. The highest incorporation rates are obtained with 3′ overhangs.

Application

Use terminal transferase to add nucleotides to the 3′-OH ends of double- or single-stranded DNA fragments, for example:
  • Tailing with dNTPs:Addition of homopolymeric tails to DNA fragments
  • Labeling of double- and single-stranded DNA and oligonucleotides with either radioactive or chemically modified nucleotides (e.g., DIG-dUTP)
3′-end Labeling with ddNTPs:
Labeling of double- and single-stranded DNA and oligonucleotides with either radioactive or chemically modified dideoxynucleotides (e.g., DIG-ddUTP)

Biochem/physiol Actions

Oligonucleotides are enzymatically labeled at their 3′ end using terminal transferase by incorporation of a single digoxigenin-labeled dideoxyuridine-triphosphate. Another way to label oligonucleotides is the addition of a longer nucleotide tail. For the generation of tailed oligonucleotide probes, deoxynucleotides triphosphates are used in a template independent reaction.

Features and Benefits

Incorporation of labeled or modified nucleotides
In addition to standard nucleotides, terminal transferase wlll add radioactive or modified (e.g., digoxigenin-, biotin-, or fluorochrome-labeled) dNTPs or ddNTPs to DNA.

Packaging

1 kit containing 3 components

Preparation Note

Working solution: Standard Tailing reaction with radioactive nucleotides
Preparation of CoCl2 working solution
Add in a sterile vial 10 μl double dist. water and 15 μl of the supplied 25 mM CoCl2 solution: Final concentration: 15 mM

Preparation of radioactive labeling mix
dATP and dTTP labeling mix: mix 1 Vol. of a 2.5 mM dATP or dTTP solution with 15 volumes of double-distilled water and 4 volumes of α-32P-dATP or α-32P-dTTP (800 Ci/mmol, approx. 30 TBq/mmol).
dGTP and dCTP labeling mix: mix 1 volume of a 2 mM dGTP or dCTP solution with 15 volumes of double-distilled water and 4 volumes of α-32P-dGTP or α-32P-dCTP (800 Ci/mmol, approx. 30 TBq/mmol)

Analysis Note

Absence of 5′ and 3′ exonucleases, endonucleases, and nicking activities tested according to the current Quality Control procedures.

Other Notes

For general laboratory use. Double-stranded DNA may have either blunt-, 3′-protruding, or 5′-protruding ends. However, 3′-protruding ends lead to the highest incorporation rates.TdT requires an oligonucleotide of at least three bases as a primer, and single-stranded DNA is tailed more efficiently than double-stranded.
One unit is the enzyme activity that incorporates 1 nMol dTMP into acid-insoluble products within 30 minutes at +37 °C under assay conditions using d(pT)6 as primer. Unit assay conditions: 200 mM Potassium cacodylate, 1 mM CoCl2, 1 mM dTTP, 0.1 OD d(pT)6, 6.25 pmol 3H dTTP in a 120 μl reaction volume.

Unit Assay: Unit assay conditions: 200 mM Potassium cacodylate, 1 mM CoCl2, 1 mM dTTP, 0.1 OD d(pT)6, 6.25 pmol 3H dTTP in a 120 μl reaction volume.

Volume Activity: 400 U/μl

Sample Materials
  • Double- or single-stranded DNA fragments
  • Double- or single-stranded oligonucleotides

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Sigma-Aldrich

338A7

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description

from Calf Thymus, recombinant, E. coli

description

from Escherichia coli BL 21/pAR 1219

description

from recombinant E.Coli

description

-

application(s)

genomic analysis

application(s)

life science and biopharma

application(s)

-

application(s)

-

form

solution

form

solution

form

solution

form

buffered aqueous solution

recombinant

expressed in E. coli

recombinant

-

recombinant

-

recombinant

-

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

2-8°C

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Cell Marque®

packaging

pkg of 24,000 U (03333574001 [400 U per reaction]), pkg of 8,000 U (03333566001 [400 U per reaction])

packaging

pkg of 1,000 U (10881767001), pkg of 5,000 U (10881775001)

packaging

pkg of 1,000 U (10674257001 [10 U/μl]), pkg of 20,000 U (10674273001 [10 U/μl]), pkg of 20,000 U (11047663001 [40 U/μl]), pkg of 5,000 U (10674265001 [10 U/μl])

packaging

vial of 0.1 mL concentrate (338A-74), vial of 0.5 mL concentrate (338A-75), bottle of 1.0 mL predilute (338A-77), vial of 1.0 mL concentrate (338A-76), bottle of 7.0 mL predilute (338A-78)

Kit Components Only

Product No.
Description
  • Terminal Transferase 400 U/μl
  • TdT Reaction Buffer 5x concentrated
  • CoCl<sub>2</sub> Solution 25 mM

signalword

Danger

wgk

WGK 3

flash_point_f

does not flash

flash_point_c

does not flash

Hazard Classifications

Acute Tox. 4 - Acute Tox. 4 Oral - Aquatic Chronic 2 - Carc. 1B Inhalation - Repr. 1B Inhalation

Storage Class

6.1D - Non-combustible acute toxic Cat.3 / toxic hazardous materials or hazardous materials causing chronic effects

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