Sanger sequencing
E561076
Sanger sequencing is a DNA sequencing method that uses chain-terminating dideoxynucleotides to determine the precise order of nucleotides in a DNA molecule.
Statements (50)
| Predicate | Object |
|---|---|
| instanceOf |
DNA sequencing method
ⓘ
molecular biology technique ⓘ |
| alsoKnownAs |
chain-termination sequencing
ⓘ
dideoxy sequencing ⓘ |
| basedOn | DNA replication ⓘ |
| commonlyUsedFor |
clinical diagnostic sequencing of specific loci
ⓘ
sequencing individual genes ⓘ small-scale sequencing projects ⓘ validating variants detected by high-throughput sequencing ⓘ |
| contributedTo |
Human Genome Project
NERFINISHED
ⓘ
sequencing of the first complete genomes ⓘ |
| determines | nucleotide sequence of DNA ⓘ |
| developedBy | Frederick Sanger NERFINISHED ⓘ |
| developedInYear | 1977 ⓘ |
| enabled | rapid sequencing of cloned DNA fragments ⓘ |
| errorTypesInclude |
base miscalls
ⓘ
uncertain base calls at low signal regions ⓘ |
| higherAccuracyThan | many next-generation sequencing methods per base ⓘ |
| limitationsInclude |
difficulty with long homopolymer regions
ⓘ
reduced quality near end of read ⓘ requirement for prior knowledge to design primers ⓘ |
| lowerThroughputThan | next-generation sequencing ⓘ |
| moreWidelyAdoptedThan | Maxam–Gilbert sequencing NERFINISHED ⓘ |
| nowOftenUses |
capillary electrophoresis
ⓘ
fluorescently labeled dideoxynucleotides ⓘ |
| originallyAnalyzedBy | polyacrylamide gel electrophoresis ⓘ |
| originallyUsed |
four separate reactions
ⓘ
radioactively labeled nucleotides ⓘ |
| output | electropherogram ⓘ |
| produces | DNA fragments of varying length ⓘ |
| readLength | approximately 500–1000 bases ⓘ |
| reliesOn | selective incorporation of chain-terminating dideoxynucleotides ⓘ |
| requires |
annealing of primer
ⓘ
clonal DNA template ⓘ denaturation of DNA ⓘ extension by DNA polymerase ⓘ |
| templateCanBe |
M13 phage DNA
ⓘ
PCR product ⓘ plasmid DNA ⓘ |
| usedIn |
genotyping of specific loci
ⓘ
molecular cloning workflows ⓘ mutation detection ⓘ |
| uses |
DNA polymerase
ⓘ
DNA template ⓘ chain-terminating nucleotides ⓘ deoxynucleotides ⓘ dideoxynucleotides ⓘ primer ⓘ |
| wasKeyTo | first-generation DNA sequencing era ⓘ |
| wasPrecededBy | Maxam–Gilbert sequencing NERFINISHED ⓘ |
Referenced by (2)
Full triples — surface form annotated when it differs from this entity's canonical label.