recombinant technology

Question 1

what is a multiple cloning site

Answer 1

part of plasmid where many restriction enzymes are added

Question 2

why are yeast artificial chromosomes difficult to work with?

Answer 2

they are large, fragile and break easily

Question 3

what are the sources of DNA

Answer 3

• genomic library

- cDNA library

Question 4

automated sequencing

Answer 4

sequences up to 1000 nucleotides at a time

1. ddNTPs labelled with fluorescent dye
2. as sequencing gel is running, a camera takes a picture of the same position over time and measures intensity
3. graph obtained of colour intensity over time
4. peaks correspond with a ddNTP

Question 5

how many base pairs does the human genome consist of?

Answer 5

3.2^9

Question 6

progressive sequencing

Answer 6

use BACs
design primers based on either end of known vector sequences reading in direction of insert
sequence part of DNA sequence
design primers based on newly known DNA sequence
repeat until sequences meet in the middle

Question 7

shotgun sequencing

Answer 7

1. make genomic plasmid library
2. sequence ends of each clone using primers based on vector
3. reconstruct original sequence based on sequences assembled and overlapping = contig

Question 8

what are methods used to sequence the human genome?

Answer 8

• progressive sequencing

- shotgun sequencing