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Separate analysis of complementary strands of restriction enzyme-digested DNA. An application of restriction fragment mass mapping by matrix-assisted laser desorption/ionization mass spectrometry

✍ Scribed by Wada, Yoshinao

Publisher: John Wiley and Sons
Year: 1998
Tongue: English
Weight: 216 KB
Volume: 33
Category: Article
ISSN: 1076-5174
DOI: 10.1002/(sici)1096-9888(199802)33:2<187::aid-jms625>3.0.co;2-a

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✦ Synopsis

Matrix-assisted laser desorption/ionization time-of-Ñight mass spectrometry (MALDI/TOF-MS) of a restriction endonuclease digest determines the molecular mass of PCR-ampliÐed DNA more easily than measurement of undigested DNA. With this method, a 664 bp region from the FAS gene could be analyzed and a two-nucleotide deletion in the L1CAM gene was detected in a restriction fragment of 105 nucleotides. Furthermore, the analysis of smaller fragments allowed separate detection of single-stranded oligonucleotides comprising individual digested fragments. This mixture analysis of restriction enzyme digests improves the resolution, sensitivity and accuracy of MALDI/TOF-MS of DNA and is thus expected to facilitate its application to genetic diagnosis.

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