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About sequencing

Revision as of 20:39, 4 October 2017 by imported>이경준

In genomics, sequencing is to determine the arrangement of DNA. Sequencing methods are outlined with three basic steps as shown below.

 

  1. Library preparation

  2. Template preparation

  3. 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