1. a) Starting with a single molecule of an mRNA template and sufficient primers

Question

1. a) Starting with a single molecule of an mRNA template and sufficient primers, enzyme and all other co-factors for successful gene specific RT-PCR, how may PCR cycles are required to have 8 molecules of products with ends defined by both primers? By this I mean that you count only double stranded molecules that begin and end at the primer binding sites but lack any other sequence.

b) I want to use RT-PCR to make a copy of the entire Liliputian gene. Indicate, the sequence of a primer of 6 nucleotides that could be used for the first strand synthesis of cDNA.

Explanation / Answer

RT PCR means reversre Transcrition Polymerase chain reaction. In this we first reverse transcribe the RNA of interest into its DNA complement through the use of reverse transcriptase. Subsequently, the newly synthesized cDNA is amplified using traditional PCR.

In case of Traditional PCR

Number of copies of DNA obtained after 'n' cycles = 2(n+1)

so, by this we get n = 3 cycles to have 8 molecules of products and

one additional cycle would be required to convert mRNA to cDNA.

so, total should be 3+1 =4

b)

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