Template‐Independent DNA Polymerases

Dongxian Yue1, Stanley Tabor2, Nicole M. Nichols1

1 New England Biolabs, Ipswich, Massachusetts, 2 Harvard Medical School, Boston, Massachusetts
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 3.6
DOI:  10.1002/0471142727.mb0306s84
Online Posting Date:  October, 2008
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Abstract

Terminal deoxynucleotidyl transferase (TdT), is a template‐independent DNA polymerase that catalyzes the incorporation of deoxynucleotides at the 3′‐hydroxyl terminus of DNA, accompanied by the release of inorganic phosphate. TdT does not require a template and will not copy one. Reaction conditions and some applications are described in this unit, including cloning DNA fragments, labeling the 3′ terminus of DNA with 32P or nonradioactive tags, synthesizing model polydeoxynucleotide homopolymers, and detecting DNA damage and apoptotic cells. Curr. Protoc. Mol. Biol. 84:3.6.1‐3.6.3. © 2008 by John Wiley & Sons, Inc.

Keywords: terminal deoxynucleotidyl transferase; TdT; DNA 3′‐end labeling; DNA tailing; TUNEL assay

     
 
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Table of Contents

  • Enzyme: Terminal Deoxynucleotidyl Transferase (Terminal Transferase or TdT)
  • Literature Cited
  • Figures
     
 
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Materials

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Figures

Videos

Literature Cited

   Deng, G.R. and Wu, R. 1983. Terminal transferase: Use in the tailing of DNA and for in vitro mutagenesis. Methods Enzymol. 100:96‐116.
   Labat‐Moleur, F., Guillermet, C., Lorimier, P., Robert, C., Lantuejoul, S., Brambilla, E., and Negoescu, A. 1998. TUNEL apoptotic cell detection in tissue sections: Critical evaluation and improvement. J. Histochem. Cytochem. 46:327‐334.
   Rohwer, F. and Azam, F. 2000. Detection of DNA damage in prokaryotes by terminal deoxyribonucleotide transferase‐mediated dUTP nick end labeling. Appl. Environ. Microbiol. 66:1001‐1006.
   Tu, C.‐P.D. and Cohen, S.N. 1980. 3′‐End labeling of DNA with [α‐32P]cordycepin‐5′‐triphosphate. Gene 10:177‐183.
   Vincent, C., Tchen, P., Cohen‐Solal, M., and Kourilsky, P. 1982. Synthesis of 8‐(2,4‐dinitrophenyl‐2,6‐aminohexyl)aminoadenosine‐5′‐triphosphate: Biological properties and potential uses. Nucleic Acids Res. 10:6787‐6796.
Key Reference
   Ratliff, R.L. 1981. Terminal deoxynucleotidyl transferase. In The Enzymes, Vol. 14A (P.D. Boyer, ed.) pp. 105‐118. Academic Press, New York.
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