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 Protocol for DNA Tailing
(with Terminal Deoxynucleotidyl Transferase)

1. Prepare the following reaction mixture:
   

   5X reaction buffer for Terminal Deoxynucleotidyl Transferase	4 µl
   DNA fragments	1 pmol of 3'-termini
   dATP or dTTP or dGTP or dCTP	130 pmol 60 pmol
   Terminal Deoxynucleotidyl Transferase	1.5 µl (30 u)
   Water, nuclease-free	to 20 µl

2. Incubate the mixture at 37°C for 15 minutes.
3. Stop the reaction by heating at 70°C for 10 minutes or by the addition of 2 µl 0.5M EDTA (pH 8.0).

Note
Under the conditions described above, 100-130 dA or dT residues, or 20-30 dC or dG residues per 3'-OH end of DNA can be added.

References

1. Deng, G.R., Wu, R., Terminal transferase: Use in the tailing of DNA and for in vitro mutagenesis, Meth. Enzymol., 100, 96-116, 1983.
2. Eschenfeldt, W.H., et al., Homopolymeric tailing, Meth. Enzymol., 152, 337-342, 1987.

Ordering Information

Terminal Deoxynucleotidyl Transferase dNTP Set M-MuLV Reverse Transcriptase RevertAid™ M-MuLV Reverse Transcriptase RevertAid™ M-MuLV Reverse Transcriptase Modified Nucleotides (molecular biology grade): Aminoallyl-dUTP  Biotin-11-dUTP Fluorescein-12-dUTP dm6ATP dm4CTP dm5CTP 0.5 M EDTA, pH 8.0 Water, nuclease-free

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Updated gegužės 09, 2007 17:26