ISSN: 1755-5191

Imaging in Medicine

Sequencing Open

 Next-generation sequencing involves three basic steps: library preparation, sequencing, and data analysis. The principle behind Next Generation Sequencing (NGS) is similar to that of Sanger sequencing, which relies on capillary electrophoresis. The genomic strand is fragmented, and the bases in each fragment are identified by emitted signals when the fragments are ligated against a template strand. Sanger methods can only sequence short pieces of DNA--about 300 to 1000 base pairs. The quality of a Sanger sequence is often not very good in the first 15 to 40 bases because that is where the primer binds. Sequence quality degrades after 700 to 900 bases. A DNA polymerase enzyme. A primer, which is a short piece of single-stranded DNA that binds to the template DNA and acts as a "starter" for the polymerase. The four DNA nucleotides (dATP, dTTP, dCTP, dGTP) The template DNA to be sequenced. PCR uses forward and reverse primers. The forward primer anneals to a complimentary site on one strand of DNA and extends toward the reverse primer. ... Sanger sequencing uses one primer instead of two. The amplification process copies one strand but not the reverse strand. Testing may increase anxiety and stress for some individuals.  

High Impact List of Articles

Relevant Topics in General Science