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Updated: Feb 11, 2026

Identification of Homologous Recombination Events in Mouse Embryonic Stem Cells Using Southern Blotting and Polymerase Chain Reaction
Published on: November 20, 2018
Validation of quantitative polymerase chain reaction with Southern blot method for telomere length analysis
Mohamad Tarik1,1, Lakshmy Ramakrishnan1,1, Harshpal S Sachdev2,2
1Department of Cardiac Biochemistry, All India Institute of Medical Science, New Delhi, India.
Aim:
Telomere length (TL) measurement by quantitative polymerase chain reaction (PCR) has been widely accepted, but limited information regarding its validation with a gold-standard technique is available.
Materials & Methods:
We measured TL by Southern blot and monochrome multiplex quantitative PCR (MMqPCR) and validated the results of TL in leukocytes of 94 participants with mean age 43.2 years, BMI 19-41 (mean 27.8 ± 4.3) kg/m2.
Results:
A significant positive correlation was observed between TL measured by MMqPCR and Southern blot assay (correlation coefficient r = +0.896, p < 0.0001). The inter- and intra-assay CVs of the MMqPCR assay were 5.3 and 4.07%, respectively.
Conclusion:
We observed that experimental discrepancies have an impact on TL analysis and there is a need to improve the optimum conditions.
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