Questions on cDNA 1. When we start making cDNA with just the eukaryotic mRNA as

Question

Questions on cDNA 1. When we start making cDNA with just the eukaryotic mRNA as a template we usually use as a primer for making the first strand of cDNA? 2. Why do we start synthesizing cDNA from the 3' end of the mRNA? 3. After you have synthesized the first strand of eDNA, what primer(s) could you use to make the second strand of cDNA? 4. When making cDNA, what general ingredients must be in the tube to synthesize the first strand of cDNA? S. If you use random hexamers to prime the second strand of your cDNA, how could you get rid of the mRNA to get single stranded first strand cDNA?

Explanation / Answer

1) Complimentary DNA is usually abbreviated as cDNA. It is formed from mRNA template by the action of enzymes reverse transcriptase and DNA polymerase. The primer that is usually used in the synthesis of first strand of cDNA is oligo-dT (18).

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