DNA tailing without terminal transferase

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DNA tailing without terminal transferase.

Tailing of DNA with terminal transferase is a technically demanding, but essential step in full-length cDNA cloning procedures*•^. It is also an essential step of a poly(dA) affinity labelling technique, which has been used to select families of partial restriction fragments on oligo(dT)cellulose^•*. In order to bypass the need for transferase in the latter technique, we made a simple modificat...

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Tailing cDNAs with terminal deoxynucleotidyl transferase in RT-PCR assays to identify ribozyme cleavage products

Polytailing a cDNA with terminal deoxynucleotidyltransferase (TdT) results in the addition of a homopolymeric sequence at its 3'-end. Here we describe the use of tailing in competitive RT-PCR assays to evaluate cleavage efficiency of ribozymes. Using a system that perfectly mimics intracellular cleavage, we were able to detect as few as 1% of cleaved moieties. Furthermore, employing primers ove...

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Measurement of DNA damage by terminal deoxynucleotidyl transferase reaction

An improved terminal deoxynucleotidyl transferasemediated dUTP-biotin nick end labeling method for the quantification of DNA damage in tissues and cultured cells was developed. Many reports have revealed that histochemistry of DNA damage can be visualized using immunohistochemistry for the terminal deoxynucleotidyl transferase reaction in tissue sections. However, few reports have described qua...

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Terminal deoxynucleotidyl transferase: the story of a misguided DNA polymerase.

Nearly every DNA polymerase characterized to date exclusively catalyzes the incorporation of mononucleotides into a growing primer using a DNA or RNA template as a guide to direct each incorporation event. There is, however, one unique DNA polymerase designated terminal deoxynucleotidyl transferase that performs DNA synthesis using only single-stranded DNA as the nucleic acid substrate. In this...

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Terminal deoxynucleotidyl transferase catalyzes the reaction of DNA phosphorylation.

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ژورنال

عنوان ژورنال: Nucleic Acids Research

سال: 1988

ISSN: 0305-1048,1362-4962

DOI: 10.1093/nar/16.15.7744