期刊
BIOTECHNIQUES
卷 60, 期 6, 页码 306-309出版社
BIOTECHNIQUES OFFICE
DOI: 10.2144/000114427
关键词
oligonucleotide; telomere; bridged nucleic acid (BNA); terminal restriction fragments (TRF)
资金
- NIH [C06 RR30414]
- National Institute on Aging [AG01228]
- CPRIT Postdoctoral Cancer Intervention and Prevention Discoveries Training Program [RP140110]
- Harold Simmons NCI Designated Comprehensive Cancer Center Support Grant [CA142543]
- Southland Financial Corporation Distinguished Chair in Geriatric Research
Telomere length in humans has been correlated with cancer and age-related diseases. The standard method to measure telomere length relies on Southern blot analysis with radioactively or nonradioactively labeled probes containing several telomeric DNA repeats. However, this approach requires relatively large amounts of genomic DNA, making it difficult to measure telomere length when a limited amount of sample is available. Here, we describe a non-radioactive labeling method that uses 3'fill-in combined with lambda exonuclease digestion to incorporate one or more digoxigenin (DIG) molecules into bridged nucleic acid (BNA)-containing oligonucleotides (ONTs). Using our method, we were able to generate probes to detect both C-and G-rich telomeric DNA strands. Compared with commercially available DIG-labeled telomere probes, probes generated using this new approach significantly enhance the sensitivity of telomere length measurements.
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