Difference between revisions of "Next Generation Sequencing"
imported>Sanzhar Aitbay |
imported>Sanzhar Aitbay |
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<p style="margin-left:40px"><img alt="" src="/ckfinder/userfiles/images/Picture2(1).png" style="height:13px; width:200px" /></p> | <p style="margin-left:40px"><img alt="" src="/ckfinder/userfiles/images/Picture2(1).png" style="height:13px; width:200px" /></p> | ||
| − | <p style="margin-left: 80px | + | <p style="margin-left:80px">Sequencing technologies include a number of methods<br /> |
that are grouped broadly as <strong>template preparation,<br /> | that are grouped broadly as <strong>template preparation,<br /> | ||
sequencing and imaging, and data analysis.</strong></p> | sequencing and imaging, and data analysis.</strong></p> | ||
<ul> | <ul> | ||
| − | <li | + | <li> |
<p><em>There are two methods used in preparing templates for NGS reactions</em>: <strong>clonally amplified templates</strong> originating from single DNA molecules, and<strong> single DNA molecule templates</strong></p> | <p><em>There are two methods used in preparing templates for NGS reactions</em>: <strong>clonally amplified templates</strong> originating from single DNA molecules, and<strong> single DNA molecule templates</strong></p> | ||
| − | |||
| − | |||
| − | |||
</li> | </li> | ||
</ul> | </ul> | ||
| − | <p> </p> | + | <p style="margin-left: 80px;"><strong>Clonally amplified templates.</strong> most imaging systems have<br /> |
| + | not been designed to detect single fluorescent events, so<br /> | ||
| + | amplified templates are required. The two most common<br /> | ||
| + | methods are <strong>emulsion PCR (emPCR)9 and solid-phase<br /> | ||
| + | amplification</strong></p> | ||
| + | |||
| + | <p style="margin-left: 80px;"><strong><img alt="" src="/ckfinder/userfiles/images/Picture3(1).png" style="height:97px; width:400px" /><u>emulsion PCR (emPCR)</u></strong></p> | ||
| + | |||
| + | <p style="margin-left: 80px;"><strong><u><img alt="" src="/ckfinder/userfiles/images/Picture4(1).png" style="height:235px; width:400px" />solid-phase amplification</u></strong></p> | ||
| + | |||
| + | <p style="margin-left: 80px;"> </p> | ||
<p style="margin-left:40px"> </p> | <p style="margin-left:40px"> </p> | ||
Revision as of 17:31, 9 December 2017
This Review focuses on commercially available technologies from Roche/454, Illumina/Solexa, Life/APG and Helicos BioSciences, the Polonator instrument and the near-term technology of Pacific Biosciences,
Here, I present a technical review of template preparation, sequencing and imaging, genome alignment and assembly, and current NGS platform performance to provide guidance on how these technologies work and how they may be applied to important biological questions.
Sequencing technologies include a number of methods
that are grouped broadly as template preparation,
sequencing and imaging, and data analysis.
-
There are two methods used in preparing templates for NGS reactions: clonally amplified templates originating from single DNA molecules, and single DNA molecule templates
Clonally amplified templates. most imaging systems have
not been designed to detect single fluorescent events, so
amplified templates are required. The two most common
methods are emulsion PCR (emPCR)9 and solid-phase
amplification
emulsion PCR (emPCR)
solid-phase amplification
