Telomere basics and why measure telomeres?
a) Uses the O’Callaghan et al, 2008 quantitative real-time PCR (qPCR) method based on Cawthon’s (2002) qPCR method to determine absolute telomere length (aTL) values.
b) As little as 200ng DNA is required for the qPCR telomere length assay.
c) An oligomer standard generates telomere lengths (aTL) in nucleotide base pair values rather than Cawthon’s relative quantification (T/S) values.
d) Samples are measured in triplicates on the same plate, and then averaged.
e) PCR amplifications are established using a a robotic pipettor to ensure maximum pipetting accuracy.
f) Real-time PCR performed using an instrument with unique rotary design for sensitive and accurate performance to reduce measurement error.
g) In addition to running a non-template control, three established reference DNA controls are also run in triplicate on each plate as positive controls
h) Plate to plate telomere length variation is normalized using the geometric mean of the reference DNA samples.
i) Results, expressed as an average telomere length (aTL), are an actual nucleotide sequence length (kb lengths) making it easier to apply to other biomarkers such as cortisol, inflammatory cytokines or other stress biomarker levels offered for testing in the BCL
NOTE: Avoid freeze/thawing to any sample as it shortens the telomere length after sample collection. Keep samples in the -80 freezer for long-term shortage. Please complete the roster and ship the samples to the Biomarker Core Lab at the beginning of the week to avoid freeze/thaw cycling during transit.
Biomarker Core lab (BCL) now offers:
Cell separation and cell isolation from blood
DNA extraction
DNA quantification using picogreen
Telomere length determination and analysis