Question

10. Explain fully (stepwise) how to generate a cDNA library. (5 pts.) What is the advantage of this type of library? (3 pts.)

11. Compare and contrast between a cosmid, phage, Ti, and YAC vector based upon initial size, maximum insert size, cell copy number, and utility. (8 pts.)

12. Compare and contrast between the techniques of SEM and TEM. (4 pts.)

13. How is it that smaller molecules move through a gel-filtration column more slowly than larger molecules, whereas in SDS PAGE the opposite is true? Explain. (5 pts.)

14. During a DNA sequencing run, a mistake was made in that only ddNTP’s were added. How would this affect the sequencing run? (5 pts.)

Answer 1

cDNA is produced from fully transcribed mRNA found in the nucleus and therefore contains only the expressed genes of an organism. Isolate and collect MRNA Reverse h transcriptare RNA DNA insert Plasmids O GROW ☺ insect plasmids Into bacteris Isolete plasmid and purify DNA sequeance ALLT A TCGTAGT GG CIGACTIA ENA WA Formation of cDNA Library CONA is created from a mature mRNA from a eukaryotic cell with use of reverse tremsuliptare. In eukaryotes, a poly-(A) taid distinguishes mRNA som tRNA and & RNA and be used as a primer site for reverse transcription. The advantage of cDNA as that: No introns; Eukaryote genes commonly contain entrans. These are removed after mRNA synthesis so CONA wnthin, no introns. This means that cDNA copy of gene can be isolated as a single instron free fragment