Terminal Transferase
ROCHE/RTT-RO - from Calf Thymus, recombinant, E. coli
Synonym: tailing enzyme
Product Type: Chemical
| application(s) | genomic analysis |
| form | solution |
| manufacturer/tradename | Roche |
| packaging | pkg of 24,000 U (03333574001 [400 U per reaction]) |
| pkg of 8,000 U (03333566001 [400 U per reaction]) | |
| Quality Level | 100  |
| recombinant | expressed in E. coli |
| storage temp. | −20°C |
| usage | sufficient for 20 reactions (03333566001) |
| sufficient for 60 reactions (03333574001) |
| 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) |
| 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. |
| 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. |
| 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. |
| 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) |
| Principle: | Oligonucleotides are enzymatically labeled at their 3′ end using terminal transferase by incorporation of a single digoxigenin-labeled dideoxyuridine-triphospha |
| Quality: | Absence of 5′ and 3′ exonucleases, endonucleases, and nicking activities tested according to the current Quality Control procedures. |
| Unit Definition: | 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 |
| Symbol |    GHS07,GHS08,GHS09 |
| Signal word | Danger |
| Hazard statements | H302 + H332 - H350i - H360F - H411 |
| Precautionary statements | P201 - P261 - P273 - P280 - P308 + P313 - P391 |
| RIDADR | UN 3316 9 |
| WGK Germany | WGK 3 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| Storage Temp. | −20°C |
| Enzyme Commission (EC) Number | 2.7.7.31 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352200 |
| Components | Terminal Transferase 400 U/μl; TdT Reaction Buffer 5x concentrated; CoCl<sub>2</sub> Solution 25 mM |