Controlled Ribonucleotide Tailing of cDNA ends (CRTC) by Terminal Deoxynucleotidyl Transferase: A New Approach in PCR-Mediated Analysis of mRNA Sequences
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چکیده
منابع مشابه
Controlled ribonucleotide tailing of cDNA ends (CRTC) by terminal deoxynucleotidyl transferase: a new approach in PCR-mediated analysis of mRNA sequences.
Controlled ribonucleotide tailing of cDNA ends (CRTC) by terminal deoxynucleotidyl transferase is a polymerase chain reaction (PCR)-mediated technique that was developed to facilitate cloning and direct sequence analysis of complete 5'-terminal unknown coding regions of rare RNA molecules. In contrast with standard tailing protocols using dNTPs as the substrate, ribo-tailing of cDNA ends is eas...
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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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Terminal deoxynucleotidyl transferase (TdT) 1 is an enzyme which has the unique property of polymerizing deoxyribonucleotides onto a primer in the absence of a template (1-4) . Chang (4) first reported that TdT is present only in the thymus among various avian and mammalian tissues examined . Her finding was extended to humans by McCaffrey et al . (5), who found that the enzyme was restricted t...
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The recently published study of peripheral blood lymphocyte subpopulations in patients with acute immunodeficiency syndrome, by Murray et al,’ may have contributed additional insight into normal T cell ontogenesis. However, the data on terminal transferase (TdT) must be interpreted with caution. It is stated that TdT levels are especially elevated in acute lymphocytic leukemia (ALL) of T cell o...
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Several methods have been developed for determining the sequence specificity of DNA damaging agents in single-copy genes in mammalian cells (10). The ligation-mediated PCR (LMPCR) technique can be used for those compounds that cleave DNA and produce a 5′-phosphate (9,14). For those compounds that form stable adducts with DNA (e.g., cisplatin), there are three different techniques that can be us...
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ژورنال
عنوان ژورنال: Nucleic Acids Research
سال: 1996
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/24.9.1789