Comparative sequence analysis (CSA): A new sequence-based method for the identification and characterization of mutations in DNA

✍ Scribed by Christopher Mattocks; Patrick Tarpey; Martin Bobrow; Joanne Whittaker

• Publisher: John Wiley and Sons
• Year: 2000
• Tongue: English
• Weight: 385 KB
• Volume: 16
• Category: Article
• ISSN: 1059-7794
• DOI: 10.1002/1098-1004(200011)16:5<437::aid-humu9>3.0.co;2-q

✦ Synopsis

Direct sequencing analysis is largely used to confirm and characterize mutations previously detected by more rapid tests. We have developed a method-Comparative Sequence Analysis (CSA)that simplifies the analysis of sequencing data facilitating its use as a first screen for mutation detection. Sequence data were split into their component electrophoretograms and the use of a size standard enabled equivalent traces from different individuals to be overlaid. This allowed simple and rapid visual analysis of the results. Using this technique in a blind study, we tested 576 samples for mutations in the Von Hippel-Lindau tumor suppresser gene, VHL. We were able to identify and characterize all 78 known mutations present within the sample set (100% sensitivity and specificity). Hum Mutat 16:437-443, 2000.