Use of capillary electrophoresis to study methylation patterns in DNA

Karl Voss; Pieter Roos; Jian Zhong Zhang; Norman J. Dovichi

doi:10.1117/12.237608

1 April 1996 Use of capillary electrophoresis to study methylation patterns in DNA

Karl Voss, Pieter Roos, Jian Zhong Zhang, Norman J. Dovichi

Proceedings Volume 2680, Ultrasensitive Biochemical Diagnostics; (1996) https://doi.org/10.1117/12.237608
Event: Photonics West '96, 1996, San Jose, CA, United States

Abstract

A four-color multiple capillary DNA sequencer is used to determine the methylation pattern of double stranded DNA. The DNA sample is treated with bisulfite under conditions that convert cytosine to uracil. Methyl-cytosine is inert under these reaction conditions. After PCR amplification, the reaction products are subjected to a four-color fluorescent Sanger sequencing reaction. The sequence is then determined by use of capillary electrophoresis. Comparison of the sequence obtained after bisulfite treatment with the original sequence reveals that certain of the Cs in the original sequence are converted to Ts. This conversion occurs only if the original C was not methylated. Those Cs that are common to both sequences were methylated in the original sequence. Methylation patterns have been implicated in aging, developmental biology, and cancer; however, there has been no simple and rapid method for determining the methylation pattern in genomic DNA. The method described in this paper is quick, simple, and accurate, and demonstrates an exciting application of capillary electrophoresis DNA sequencing.

Citation Download Citation

Karl Voss, Pieter Roos, Jian Zhong Zhang, and Norman J. Dovichi "Use of capillary electrophoresis to study methylation patterns in DNA", Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); https://doi.org/10.1117/12.237608

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