Abstract: This invention provides methods and materials for measuring telomere abundance from chromosomes in a sample having telomeres within a pre-determined length range, e.g., short telomeres up to a certain length. The methods can involve a first step of performing a time-limited extension reaction calibrated to produce extension products from a double-stranded chromosomal DNA template of no more than a defined length, and a second step of amplifying, from the extension products, sequences bounded by a sub-telomeric sequence and the anchor sequence, to produce a length-limited telomere sequence product. The abundance of telomeric sequences in this product can be measured, and the measures can be correlated to a variety of indices.

Abstract: Disclosed are methods and compositions for determining the average length or abundance of a first target nucleic by calculating the abundance of a first target nucleic acid (T) relative to the average abundance (S) of a second and a third target nucleic acid, in a single well using a separate detection label for each target nucleic acid. In various aspects, the first target nucleic acid is a telomere. In exemplary aspects, the disclosed methods and compositions can be used to determine the average telomere length in a biological sample. The average telomere length determined using the disclosed methods and compositions can be correlated to a variety of clinically important conditions and indices. This abstract is intended as a scanning tool for purposes of searching in the particular art and is not intended to be limiting of the present invention.