Asked Nov 12, 2019

What is Sanger sequencing? Why do we use ddNTP? How to read a DNA sequence gel? c. What is a cDNA seq (RNA seq)? d. What is the main difference between a genomic and a transcriptome study?


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Sanger sequencing – Developed by Frederick Sanger is a method by which the sequence of DNA can be determined. It is done by incorporating chain-terminating dideoxynucleotide triphosphates (ddNTPs) by DNA polymerase in in-vitro conditions.

The Sanger dideoxy method of DNA sequencing requires a single-stranded DNA template, a DNA primer, a DNA polymerase, normal deoxynucleotide triphosphates (dNTPs), and modified di-deoxynucleotide triphosphates (ddNTPs).

Step 3

Modified di-deoxynucleotide triphosphates (ddNTPs) are used along with the normal deoxynucleotide triphosphates (dNTPs)-  it is because the modified ddNTPs terminate DNA strand elongation. These chain-terminating nucleotides lack a 3'-OH group required for the formation of a&nbs...

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