Alternative lengthening of telomeres (ALT) is one of the two known telomere length maintenance mech- anisms that are essential for the unlimited prolifer- ation potential of cancer cells. Existing methods for detecting ALT in tumors require substantial amounts of tumor material and are labor intensive, making it difficult to study prevalence and prognos- tic significance of ALT in large tumor cohorts. Here, we present a novel strategy utilizing telomere quan- titative PCR to diagnose ALT. The protocol is more rapid than conventional methods and scrutinizes two distinct characteristics of ALT cells concur- rently: long telomeres and the presence of C-circles (partially double-stranded circles of telo- meric C-strand DNA). Requiring only 30ng of genomic DNA, this protocol will facilitate large- scale studies of ALT in tumors and can be readily adopted by clinical laboratories.
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