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Describe how dideoxynucleotides are used for sequencing DNA.

Describe how dideoxynucleotides are used for sequencing DNA.

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dideoxynucleotides are used for sequencing DNA (ddNTPs such as ddATP, ddCTP, ddTTP and ddGTP)

Sanger's method utilizes ddNTPs, DNA pol, single stranded DNA template and RNA primer. It actually utilized for chain termination.

ddNTPs are specially designed nucleotides have H attached to 3' carbon. This method causes chain termination as ddNTPs does not form the phosphodiester bond with the next deoxynucleotide. Here ddNTPs are fluorescently or radiolabeled.

In 4 samples of reaction, different(separately) ddNTPs are added along with normal dNTPs. This is followed by DNA extension by bound primer and then DNA fragments get heat denatured separated by size by gel electrophoresis. The DNA bands then visualized by autoradiography or by UV and the DNA sequence can easily be read off X-ray film or gel image.

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