You performed one of the four chain-termination reactions using ddATP, dCTP, dGT

Question

You performed one of the four chain-termination reactions using ddATP, dCTP, dGTP, and dTTP in the reaction? Your sequencing did not work. Please explain why. On the get below, you observe the following upon gel electrophoresis of the four-individual chain-termination sequencing reactions (ddA, ddC, ddG, and ddT indicate the chain-terminating nucleotide). Explain the observation, and what is the sequence of the template strand (5' rightarrow 3' left-to-rights with the ends labeled)? You have also performed a pyrosequencing reaction on the DNA. the order of addition of deoxynucleotide triphosphates is (1) dATP, (2) dCTP, (3) dGTP, and (4) dTTP, and this pattern is repeated. the template DNA sequence is 5'-ATTGTCGGTTGATTA-3' You forgot to add sulfurylase in the first reaction (following the first dATP addition), but remembered to add it for all subsequent reactions. What will be the consequence of this mistake? Explain. You forgot to add apyrase following the second round of dTTP addition [i.e., (the second time dTTP was added). What will be the consequence of this mistake? Explain.

Explanation / Answer

1b: In the absence of dNTPS, if DDNTP enters the chain will be terminated. Here in the well dATP is completely absent. Wheneever the temple is composed with T, then the ddATP will enter and immediately terminates the chain. Hence your reactions will not work.

ACCORDING TO CHEGG GUIDELINES WE HAVE TO ANSWER ONE QUESTION AT A TIME, POST THE REST AS SEPERATE QEUSTIONS. THEN I CAN HELP YOU.

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