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Describe the purpose of adding both dNTPs and ddNTPs to the reaction mix when conducting dideoxy...

Describe the purpose of adding both dNTPs and ddNTPs to the reaction mix when conducting dideoxy chain-termination DNA sequencing. Why would one want a greater abundance of dNTPs in the reaction mix than ddNTPs?

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Answer #1

ddNTP are the chain terminating nucleotide in DNA sequencing.

ddNTP lack free 3'OH which is require for further addition of nucleotide. Whenever the chain encounter the ddNTP it get terminated.

Through this we can identify the DNA sequence of the chain.

Short fragment Indicate first nucleotide and larger fragment indicate nucleotide present in the end.

dNTP are require in high amount than ddNTP because if ddNTP is present in high concentration the chain elongation will not occur and error occur while analysing the DNA sequence.

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