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TRAP-LIG, a modified telomere repeat amplification protocol assay to quantitate telomerase inhibition by small

Julie Reed1, Mekala Gunaratnam, Monica Beltran

  • 1CRUK Biomolecular Structure Group, School of Pharmacy, University of London, London WC1N 1AX, UK.

Analytical Biochemistry
|June 7, 2008
PubMed

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In eukaryotic DNA replication, a single-stranded DNA fragment remains at the end of a chromosome after the removal of the final primer. This section of DNA cannot be replicated in the same manner as the rest of the strand because there is no 3’ end to which the newly synthesized DNA can attach. This non-replicated fragment results in gradual loss of the chromosomal DNA during each cell duplication. Additionally, it can induce a DNA damage response by enzymes that recognize single-stranded DNA.
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Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
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