AAT Bioquest

What are the differences between Sanger sequencing and PCR?

Posted March 25, 2022


Sanger sequencing and PCR are two laboratory techniques that use the same starting materials. While both processes may be used in conjunction with each other, where PCR is first used to create many copies of the DNA to be sequenced, these two are distinctly different processes and neither one can replace the other.

Basis of differentiationSanger SequencingPCR
DefinitionIs the process of determining the precise nucleotide sequence in a given DNA fragmentIs the process of amplifying DNA fragments creating many copies of the DNA fragment of interest
AimTo determine the correct sequences of the bases in DNA for medical, research or criminal applicationsTo have sufficient quantity of DNA for the next process, which is sequencing
Need for ddNTPs or dNTPsRequires ddNTPs to terminate strand formationRequires dNTPs
PrimersHas only one primer reading the sequence in one direction onlyHas two primers facing each other
Additional resources

Comparison of three human papillomavirus DNA detection methods: Next generation sequencing, multiplex-PCR and nested-PCR followed by Sanger based sequencing

MagaDye™ 535-ddGTP