mRNA 5\" mRNA template is annealed to synthetic oligonucdleotide (oligo-dT) prim

Question

mRNA 5" mRNA template is annealed to synthetic oligonucdleotide (oligo-dT) primer 5 Reverse transcriptase and NTPs yield a complementary DNA strand mRNA-DNA hybrid 3" mRNA is degraded with akali 3" To prime synthesis of a second strand, an oligonucleotide of known sequence is often ligated to the 3' end of the cDNA 5' 3' DNA polymerase I and dNTPs extend the primer to yield Duplex DNA 5' 3" FIGURE 9-14 Building a cDNA library from mRNA. A cell's total mRNA content includes transcripts from thousands of genes, and the cDNAs generated from this mRNA are correspondingly heterogeneous. Reverse transcriptase can synthesize DNA on an RNA or a DNA template (see Fig. 26-32). To prime the synthesis of a second DNA strand, oligonucle- otides of known sequence are ligated to the 3" end of the first strand, and the double-stranded cDNA so produced is cloned into a plasmid

Explanation / Answer

1(d) . cDNA means -complementary DNA,because the cDNA produced are specifically complementary to mRNAs.

2, (b) the total RNA obtained from the lysed cells.

3. (c) almost all eukaryotic mRNAs have polyA tails at their 3` end

4. (c) the name "reverse" transcriptase indicates that the direction of information flow is reversed i.e, from RNA to DNA.

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