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April 22, 2021, 09:42:25 am

Author Topic: Sanger sequencing  (Read 653 times)  Share 

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Sanger sequencing
« on: October 07, 2019, 12:25:13 pm »
Hey can someone explain to me in baby language what Sanger sequencing is lol

mashal hameed

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Re: Sanger sequencing
« Reply #1 on: March 20, 2020, 02:41:57 pm »
a bit late but incase anybody else wanted to know... please correct me if i'm wrong
- DNA is isolated and replicated to there are lots of copies using polymerase chain reaction (PCR)
- DNA is heated and unwound
- A primer is annealed to the end of the DNA
- The primed DNA is divided into 4 reaction vessels (PCR was done at the start so there are lots of copies)
- dNTP (A,T,G,C) added to each
- specially modified ddNTP'S are added (special base) only one type to each (types are a,g,c,t so one vessel has a modifed a base, another a modified g base etc)
- polymerase attaches bases to template strand until the special base is paired, it terminates the sequence.
- because of termination, DNA fragments of different lengths are formed
- electrophoresis gel is used to sequence DNA
- current of electricity, makes DNA move and bands appear
* short pieces move further
- sequence read from the bottom up
results in the complementary sequence of the DNA sample