The University of Münster, Germany, used Ion Torrent sequencing technology to complete the rapid sequencing of the whole genome sequence of a new type of E. coli isolated from the patient within 62 hours. (EAEC) and Enterohaemorrhagic Escherichia coli (EHEC), a novel mixed E. coli strain. It also makes up for the shortcomings of the rapid inspection and quarantine of the next-generation sequencing technology.
Ion Torrent is the latest generation of sequencing technology. Its advent has brought exciting progress to the application of sequencing technology. Its core technology is to use semiconductor technology to establish a direct connection between chemical and digital information. DNA strands are fixed on the microspheres in the microwells of the semiconductor chip, and then ACGT is sequentially incorporated. As each base is incorporated, hydrogen ions are released, which can be detected as they pass through the bottom of each hole, and the base is interpreted in real time by detecting H +. Compared with other sequencing technologies, the sequencing system using this technology is simpler, faster, and more flexible.
Small genome sequencing DNA amplicon sequencing targeted capture sequencing
· Microbial genome
· Mitochondria
· Design specific primers for the chloroplast genome, amplify the target region-the specific genomic region of interest (thousands of amplifications or long fragments), and then perform high-throughput sequencing. Design specific capture probes to enrich target regions (specific genomic regions of interest) and then perform high-throughput sequencing.
1. Mitochondria: the difference is large, about 16Kb for mammals, and about 100 Kb for the smallest plant mitochondria
2. Chloroplast: large difference, 120 ~ 180 Kb
3. Mycoplasma pneumoniae: M.pneumoniae ~ 800 Kb
4. Streptococcus pneumoniae: S.pneumoniae ~ 2 Mb 5. Mycobacterium tuberculosis: M.tuberculosis ~ 4 Mb
6. E. coli: E. coli O157: H7 ~ 5 Mb
7. Saccharomyces cerevisiae: S.cerevisiae ~ 12 Mb
8. Cellulomycetes: S. cellulosum ~ 13 Mb (the largest known prokaryotic genome)
Mellmann A et al, Prospective genomic characterization of the German enterohemorrhagic Escherichia coli O104: H4 outbreak by rapid next generation sequencing technology, PLoS One, 2011; 6 (7): e22751.
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