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Nat Protoc. 2010 Sep;5(9):1596-607. doi: 10.1038/nprot.2010.124. Epub 2010 Sep 2.

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

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1
The Center for Human Development and Aging, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, New Jersey, USA. kimurama@umdnj.edu

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, DNA is 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 DNA sample, which are useful in answering questions beyond those focusing on the mean length of telomeres in a given sample. A skilled technician can measure TRF length in ∼130 samples per week.

PMID:
21085125
DOI:
10.1038/nprot.2010.124
[Indexed for MEDLINE]
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