MALDI-TOF based mutation detection using tagged in vitro synthesized peptides
作者： Alex M. Garvin , Kenneth C. Parker , Larry Haff
DOI: 10.1038/72013
关键词:
摘要: Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) is a powerful method to quickly and accurately determine the masses of peptides1. Most genetic analyses, however, begin with PCR amplification test sequence generate DNA, which more difficult than peptides analyze by MALDI-TOF2,3. We describe that produces product any continuous region coding can then be used encode an N-terminally tagged peptide in coupled vitro transcription/translation reaction. The purified using tag, its measured MALDI-TOF. Truncations amino acid substitutions coded for breast cancer susceptibility gene BRCA1 were readily identified this method. process multiplexed amenable automation, providing efficient, high-throughput means mutation discovery profiling.
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