Terminal Transferase, recombinant
from calf thymus, expressed in E. coli, solution
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Terminal Transferase, recombinant material number and pack size: Material Number Pack Size 03289869103 custom fill
- Terminal Transferase catalyzes the addition of deoxy- and dideoxynucleoside triphosphates to the 3'-OH ends of double- and single-stranded DNA fragments and oligonucleotides. 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 facilitates optimal tailing of all types of double-stranded DNA ends: blunt ended, with 3' overhang, or with 5' overhang.
- Use Terminal Transferase to add homopolymer tails to DNA fragments in cloning experiments, such as addition of overhanging ends onto cDNAs for easier cloning and labeling of 3' ends of double- and single-stranded DNA (e.g., oligonucleotides) with radioactive labeled nucleotides or nucleotides labeled with haptens, e.g., digoxigenin or biotin.
The enzyme catalyzes a template-independent addition of dNTPs or of a single ddNTP to the 3'-OH ends of double- or single-stranded DNA.
It accepts radioactively labeled nucleotides and nucleotides labeled with haptens, such as digoxigenin or biotin.
For activity, the enzyme requires the presence of divalent metal ions, preferably Co2+. Appearance: Clear, colorless solution
Storage buffer: Potassium phosphate, 60 mmol/L; KCl, 150 mmol/L; 2-mercaptoethanol, 1 mmol/L; Triton X-100, 0.5% (v/v); glycerol, 50% (v/v); pH approximately 7.2 at +4°C
Volume activity (Co2+): ≥400x103 U/mL
Specific activity (Co2+): ≥200x103 U/mg
Unit definition: One unit is the enzyme activity that leads to an incorporation of 1 nmol dTMP into acid insoluble products within 30 minutes at +37°C under assay conditions (cacodylate, 200 mmol/L; Co2+, 1 mmol/L) using d(pT)6 as primer.
Unspecific endonucleases (MWM II DNA): Not detectable in up to 400 U after 4 hours incubation at +37°C.
Nicking activity (pBR322 DNA): Not detectable in up to 400 U after 4 hours incubation at +37°C.
Exonucleases (3H-DNA): Not detectable in up to 1200 U.
Function test (tailing reaction on a 30-mer oligonucleotide): Corresponds to specification
Stability: At -15 to -25°C within specification range for 24 months.