Transcription of Next-Generation DNA Sequencing Methods
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Next-Generation DNA Sequencing Methods
ANRV353-GG09-20 ARI25 July 200814:57 Next-Generation DNAS equencing MethodsElaine R. MardisDepartments of Genetics and Molecular Microbiology and Genome Sequencing Center,Washington University School of Medicine, St. Louis MO 63108; email: Rev. Genomics Hum. Genet. :387 402 First published online as a Review in Advance onJune 24, 2008 TheAnnual Review of Genomics and Human Geneticsis online at article s 2008 by Annual rights reserved1527-8204/08/0922-0387$ Wordsmassively parallel Sequencing , Sequencing -by-synthesis, resequencingAbstractRecent scientific discoveries that resulted from the application of Next-Generation DNA Sequencing technologies highlight the striking impactof these massively parallel platforms on genetics. These new meth-ods have expanded previously focused readouts from a variety of DNApreparation protocols to a genome-wide scale and have fine-tuned theirresolution to single base precision. The Sequencing of RNA also hastransitioned and now includes full-length cDNA analyses, serial analysisof gene expression (SAGE)-based Methods , and noncoding RNA dis-covery.
lapping set or tiling path through each human chromosome as determined by physical map-ping(31).InBAC-basedsequencing,eachBAC clone is amplified in bacterial culture, isolated inlargequantities,andshearedtoproducesize-selected pieces of approximately 2−3 kb. These pieces are subcloned into plasmid vectors, am-
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