BACKGROUNDNext generation sequencing (NGS) expertise has revolutionized genomic and genetic analysis.
The tempo of change on this space is fast with three main new sequencing platforms having been launched in 2011: Ion Torrent’s PGM, Pacific Biosciences’ RS and the Illumina MiSeq. Here we examine the outcomes obtained with these platforms to the efficiency of the Illumina HiSeq, the present market chief.
In order to check these platforms, and get ample protection depth to permit significant evaluation, we have now sequenced a set of four microbial genomes with imply GC content material starting from 19.three to 67.7%.
Together, these characterize a complete vary of genome content material. Here we report our evaluation of that sequence knowledge in phrases of protection distribution, bias, GC distribution, variant detection and accuracy.
RESULTSSequence generated by Ion Torrent, MiSeq and Pacific Biosciences applied sciences shows close to good protection behaviour on GC-rich, impartial and reasonably AT-rich genomes, however a profound bias was noticed upon sequencing the extraordinarily AT-rich genome of Plasmodium falciparum on the PGM, leading to no protection for roughly 30% of the genome.
We analysed the flexibility to name variants from every platform and discovered that we may name barely extra variants from Ion Torrent knowledge in comparison with MiSeq knowledge, however on the expense of the next false constructive fee. Variant calling from Pacific Biosciences knowledge was attainable however larger protection depth was required.
Context particular errors have been noticed in each PGM and MiSeq knowledge, however not in that from the Pacific Biosciences platform.CONCLUSIONSAll three quick turnaround sequencers evaluated right here have been capable of generate usable sequence. However there are key variations between the standard of that knowledge and the functions it’s going to help.
Compaarison of next-generation sequencing techniques.
With quick improvement and vast functions of next-generation sequencing (NGS) applied sciences, genomic sequence info is inside attain to assist the achievement of targets to decode life mysteries, make higher crops, detect pathogens, and enhance life qualities.
NGS techniques are sometimes represented by SOLiD/Ion Torrent PGM from Life Sciences, Genome Analyzer/HiSeq 2000/MiSeq from Illumina, and GS FLX Titanium/GS Junior from Roche.
Beijing Genomics Institute (BGI), which possesses the world’s largest sequencing capability, has a number of NGS techniques together with 137 HiSeq 2000, 27 SOLiD, one Ion Torrent PGM, one MiSeq, and one 454 sequencer. We have gathered intensive expertise in pattern dealing with, sequencing, and bioinformatics evaluation.
In this paper, applied sciences of these techniques are reviewed, and first-hand knowledge from intensive expertise is summarized and analyzed to debate the benefits and specifics related to every sequencing system. At final, functions of NGS are summarized.