Telomeric repeat amplification, without shortening or lengthening of the telomerase products: a method to analyze the processivity of telomerase enzyme.
The Telomeric Repeat Amplification Protocol (TRAP) and its modified versions (including ours, TP-TRAP) change the size and/or the ratio of the telomerase products in the amplification stage of the assay. Based on our recently published method we developed a new TRAP. This method ensures that the number of telomeric repeats present in the original telomerase products does not change on PCR amplification. The usefulness of the method was proved with amplification of chemically synthesized telomerase products and a newly designed telomerase substrate oligonucleotide. This is the first report in which the PCR products directly reflect the size distribution of telomerase products generated by the enzyme.
Szatmari
I
I
Department of Biochemistry and Molecular Biology, Medical and Health Science Center, University of Debrecen, H-4012 Debrecen, Hungary.
Aradi
J
J
D002460:Cell Line; D017931:DNA Primers / Q000138:chemical synthesis; D005784:Gene Amplification; D018922:HL-60 Cells; D006367:HeLa Cells; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods; D011499:Protein Processing, Post-Translational; D012091:Repetitive Sequences, Nucleic Acid; D013379:Substrate Specificity; D019098:Telomerase / Q000235:genetics / Q000378:metabolism*; D016615:Telomere / Q000201:enzymology* / Q000235:genetics*; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D019098:Telomerase