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Principles of Sanger sequencing technique

5 years ago in Data Analysis By Subba Robin

Data analysis

I want to use Sanger’s sequencing method for the purpose of gene sequencing. Please explain the principle and procedure of this technique.

All Answers (4 Answers In All)

By Novi Tulipe Answered 5 years ago

The principle behind Sanger sequence technique are as follows: 1.A DNA primer must be attached to the template strand by hybridisation. Deoxynucleosides triphosphates must be sequentially added to the primer strand. 2.The primer must be designed for the known sequences at the end of the template strand. 3.M13 sequences should be generally attached and the primer of M13 is made. 4.The reaction mixture containing dideoxynucleoside triphosphate along with usual dNTPs. 5.During replication if ddNTPs is included instead of usual dNTPs, the growing DNA strand then the replication stops at that nucleotide 6.The ddNTP is analogue of dNTP 7.ddNTP of dNTP ends chain at its respective site.

By Pragya Answered 5 years ago

In this technique, firstly you have to prepare template. Next generate nested set of fragments. This is followed by electrolysis and gel reading. For detailed information, visit https://www.youtube.com/watch?v=FvHRio1yyhQ, https://www.khanacademy.org/science/high-school-biology/hs-molecular-genetics/hs-biotechnology/a/dna-sequencing

By Preetham M Answered 5 years ago

https://www.sciencedirect.com/topics/neuroscience/sanger-sequencing this is the best site I have ever come across that gives an explanation of Sanger sequencing technique. Kindly go through it.

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