Measurement of telomere length by the southern blot analysis of terminal restriction fragment lengths

Masayuki Kimura, Rivka C. Stone, Steven C. Hunt, Joan Skurnick, Xiaobin Lu, Xiaojian Cao, Calvin B. Harley, Abraham Aviv

Research output: Contribution to journalArticlepeer-review

295 Scopus citations

Abstract

In this protocol we describe a method to obtain telomere length parameters using southern blots of terminal restriction fragments (trFs). We use this approach primarily for epidemiological studies that examine leukocyte telomere length. However, the method can be adapted for telomere length measurements in other cells whose telomere lengths are within its detection boundaries. after extraction, Dnais inspected for integrity, digested, resolved by gel electrophoresis, transferred to a membrane, hybridized with labeled probes and exposed to X-ray film using chemiluminescence. although precise and highly accurate, the method requires a considerable amount of Dna(3 µg per sample) and it measures both the canonical and noncanonical components of telomeres. the method also provides parameters of telomere length distribution in each Dnasample, which are useful in answering questions beyond those focusing on the mean length of telomeres in a given sample. askilled technician can measure trF length in ˜130 samples per week.

Original languageEnglish (US)
Pages (from-to)1596-1607
Number of pages12
JournalNature protocols
Volume5
Issue number9
DOIs
StatePublished - Sep 2 2010
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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