ΩqPCR determines absolute telomere length in kb units from single cells. Accuracy and precision of ΩqPCR were assessed using 800 bp and 1600 bp synthetic telomeres inserted into plasmids, which were measured to be 819 ± 19.6 and 1590 ± 42.3 bp, respectively. This is the first telomere length measuring method verified in this way. The approach uses Ω-probes, a DNA strand containing sequence information that enables: (i) hybridization with the telomere via the 3′ and 5′ ends that become opposed; (ii) ligation of the hybridized probes to circularize the Ω-probes and (iii) circularized-dependent qPCR due to sequence information for a forward primer, and for a reverse primer binding site, and qPCR hydrolysis probe binding. Read through of the polymerase during qPCR occurs only in circularized Ω-probes, which quantifies their number that is directly proportional to telomere length. When used in concert with information about the cell cycle stage from a single-copy gene, and ploidy, the MTL of single cells measured by ΩqPCR was consistent with that obtained from large sample sizes by TRF.

中文翻译：

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ΩqPCR 以碱基对为单位测量单细胞的端粒长度

ΩqPCR 确定单个细胞中以 kb 为单位的绝对端粒长度。使用插入质粒的 800 bp 和 1600 bp 合成端粒评估 ΩqPCR 的准确性和精密度，分别测量为 819 ± 19.6 和 1590 ± 42.3 bp。这是第一个以这种方式验证的端粒长度测量方法。该方法使用 Ω-probes，一种包含序列信息的 DNA 链，能够： (i) 通过 3' 和 5' 末端与端粒杂交；(ii) 由于正向引物和反向引物结合位点和 qPCR 水解探针结合的序列信息，杂交探针的连接以环化 Ω-探针和 (iii) 环化依赖性 qPCR。qPCR 期间聚合酶的通读仅发生在环状 Ω 探针中，它量化了与端粒长度成正比的数量。当与来自单拷贝基因的细胞周期阶段和倍性信息一起使用时，通过 ΩqPCR 测量的单细胞的 MTL 与通过 TRF 从大样本量获得的 MTL 一致。