Emulsion PCR isolates individual DNA molecules along with primer-coated beads in aqueous droplets within an oil phase.Modify 3’ ends of clones attach beads to surface.Attach library molecules to beads amplify library by emulsion PCR.Prepare either single or ‘mate-pair’ library from DNA fragments. ABI acquired the SOLiD technology from Agencourt in 2006.George Church licensed his ‘polony’ technique to Agencourt Personal Genomics.SOLiDSequencing by Oligonucleotide Ligation and Detection Chromatin Immunoprecipitation (ChIP-Seq).Sequencing representation biases, especially at beginning.Most widespread technology so that comparisons seemeasier.Different linkers and primers are attached to each end Paired-End Illumina Method Paired-end reads are easy on Illumina because the clusters are generated by ligated linkers. Illumina (Solexa) Genome Analyzer and Flow Cell new long reads from Illumina are competitive.Used to be best for metagenomics by random sequencing.De novo small genome (prokaryote or small eukaryote genome) sequencing.CON Cost is relatively high Frequent errors in runs of bases Frequent G-A transitions.PRO Long reads are uniquely identifiable Relatively quick ~20 hours total.Mapping of chromatin interactions (HiC) (courtesy Elemento lab)įounded by Jonathan Rothberg as a secret project (code-named ‘454’) within CuraGen Paired-end reads on opposite strands can be made by most technologiesĭNA methylation profiling mC C C U After PCR C C U T PCR+Seq.Generating many thousands or millions of short (30 to 1,000 base) sequences by sequencing parts of longer (200+ base) DNA fragments.Roche, Illumina, SOLiD, Ion Torrent, etc….What technologies are now available and coming?.What can we do with next-generation sequencing?.High-Throughput Sequencing Advanced Genomic Data Analysis BIOS 691-804, 2012 Mark Reimers
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