Difference between revisions of "Sanger sequencing"
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| − | < | + | <span style="font-size:22px;">'''1<sup>st</sup> sequencing – Sanger sequencing'''</span> |
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| − | + | *'''Sanger sequencing by capillary electrophoresis is the gold-standard DNA sequencing technique.''' | |
| + | *'''Based on the selective incorporation of chain-terminating ddNTP by DNA polymerase during in vitro DNA replication.''' | ||
| + | *'''requires a single-stranded DNA template, a DNA primer, a DNA polymerase, normal deoxynucleosidetriphosphates (dNTPs), and modified di-deoxynucleosidetriphosphates (ddNTPs)''' | ||
| + | *'''resulted DNA fragments are heat denatured and separated by size using gel electrophoresis''' | ||
| + | *'''DNA polymerases copy single-stranded DNA templates by adding nucleotides to a growing chain. Chain elongation occurs at the 3' end of a primer, an oligonucleotide that anneals to the template. The deoxynucleotides added to the extension product are selected by base-pair matching to the template.''' | ||
| + | *'''Thermal cycling -> purification -> capillary electrophoresis''' | ||
| − | + | [http://media.invitrogen.com.edgesuite.net/ab/applications-technologies/pharma-biotherapeutics/DNA_sequencing.swf '''http://media.invitrogen.com.edgesuite.net/ab/applications-te'''chnologies/pharma-biotherapeutics/DNA_sequencing.swf] | |
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| + | [https://en.wikipedia.org/wiki/Sanger_sequencing https://en.wikipedia.org/wiki/Sanger_sequencing] | ||
Latest revision as of 10:01, 7 March 2024
1st sequencing – Sanger sequencing
- Sanger sequencing by capillary electrophoresis is the gold-standard DNA sequencing technique.
- Based on the selective incorporation of chain-terminating ddNTP by DNA polymerase during in vitro DNA replication.
- requires a single-stranded DNA template, a DNA primer, a DNA polymerase, normal deoxynucleosidetriphosphates (dNTPs), and modified di-deoxynucleosidetriphosphates (ddNTPs)
- resulted DNA fragments are heat denatured and separated by size using gel electrophoresis
- DNA polymerases copy single-stranded DNA templates by adding nucleotides to a growing chain. Chain elongation occurs at the 3' end of a primer, an oligonucleotide that anneals to the template. The deoxynucleotides added to the extension product are selected by base-pair matching to the template.
- Thermal cycling -> purification -> capillary electrophoresis
