Difference between revisions of "About sequencing"
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| − | <p><span style="color:#000000">In genomics, sequencing is to determine the arrangement </span><span style="color:#000000">of DNA. </span>Sequencing | + | <p><span style="color:#000000">In genomics, sequencing is to determine the arrangement </span><span style="color:#000000">of DNA. </span>Sequencing methods are outlined with three basic steps as shown below.</p> |
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<p><span style="color:#000000">A genomic library is a collection of the total genomic DNA</span><span style="color:#000000"> from a single organism</span><span style="color:#000000">. The DNA is stored in a population of identical vectors,</span><span style="color:#000000"> each containing a different insert </span><span style="color:#000000">of DNA. In order to construct a genomic library, the organism's DNA is extracted </span><span style="color:#000000">from cells </span><span style="color:#000000">and then digested with a restriction enzyme to cut the DNA into fragments of a specific size. (above definition is by https://en.wikipedia.org/wiki/Genomic_library)</span></p> | <p><span style="color:#000000">A genomic library is a collection of the total genomic DNA</span><span style="color:#000000"> from a single organism</span><span style="color:#000000">. The DNA is stored in a population of identical vectors,</span><span style="color:#000000"> each containing a different insert </span><span style="color:#000000">of DNA. In order to construct a genomic library, the organism's DNA is extracted </span><span style="color:#000000">from cells </span><span style="color:#000000">and then digested with a restriction enzyme to cut the DNA into fragments of a specific size. (above definition is by https://en.wikipedia.org/wiki/Genomic_library)</span></p> | ||
| − | <p | + | <p>Fragments made by restriction enzyme undergo additional process, such as ligation of adaptor and primer, which let them be able to function in next phase.</p> |
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<h2>emPCR</h2> | <h2>emPCR</h2> | ||
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| + | <p><img alt="" src="/ckfinder/userfiles/images/emPCR%2C%20Solid-phase%20amplification%20(2).png" style="height:183px; width:731px" /></p> | ||
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| + | <p> </p> | ||
<p> </p> | <p> </p> | ||
<h2>Solid-phase</h2> | <h2>Solid-phase</h2> | ||
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| + | <p><img alt="" src="/ckfinder/userfiles/images/emPCR%2C%20Solid-phase%20amplification%20(3).png" style="height:285px; width:478px" /></p> | ||
<p> </p> | <p> </p> | ||
Revision as of 20:39, 4 October 2017
In genomics, sequencing is to determine the arrangement of DNA. Sequencing methods are outlined with three basic steps as shown below.
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Library preparation
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Template preparation
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Sequencing and imaging
Contents
Library preparation
A genomic library is a collection of the total genomic DNA from a single organism. The DNA is stored in a population of identical vectors, each containing a different insert of DNA. In order to construct a genomic library, the organism's DNA is extracted from cells and then digested with a restriction enzyme to cut the DNA into fragments of a specific size. (above definition is by https://en.wikipedia.org/wiki/Genomic_library)
Fragments made by restriction enzyme undergo additional process, such as ligation of adaptor and primer, which let them be able to function in next phase.
As shown as 'Good fragment', fragments are expected to have adaptor(blue colored zone in the above picture) allowing fragments to attached to emPCR bead and solid phase which function for the template preparation .
Template preparation
Template preparation is to generate and amplify the template DNA, that is the main target of sequencing. there are several different ways in preparing the template DNA as much as diverse secuencing methods exist.
emPCR
Solid-phase
Sequencing and imaging
