Telomerase Activity in Immortalized Cells
1. Telomerase is a ribonucleoprotein complex that consists of telomerase RNA (which contains several tandem hexanucleotide repeats complementary to the telomere repeats in the substrate) and a reverse transcriptase. This complex is able to synthesize telomeric repetitive DNA sequences at the end of linear DNA molecules. Telomerase RNA binds to the DNA strand, and complementary dNTPs are incorporated into newly synthesized, extended DNA strands by reverse transcriptase. Thus, dNTPs must also have been present during step 1 of the TRAP assay (step 1).
2. The PCR reaction mixture also contained a heat-resistant DNA polymerase (Taq polymerase).
3. Sample 3 contained cell extract isolated from immortal cells. In the presence of the telomerase, products...