Topic 10: Recombinant DNA

Question 1

Plasmids

Answer 1

covalently closed circular extrachromosomal DNA (ccccDNA)
<~10kbp

Question 2

Viruses/phage

Answer 2

~15kbp

Question 3

Cosmids

Answer 3

combination of viruses and plasmids
- concatonate, clone into plasmid, cut + circularize to look like a plasmid
- can take bigger inserts (35-40kbp)

Question 4

BAC/PAC/YAC

Answer 4

large insert vectors
(Bacterial Artificial Chromosomes, P1, Yeast)
- up to 1 000 000 kbps

Question 5

cDNA libraries

Answer 5

collections of DNA copies of transcribed sequences (mRNA) from a specific tissue

Question 6

Genomic libraries

Answer 6

collections of DNA which together cover the entire genome of an organism

Question 7

Tm

Answer 7

Primer’s melting point + therefore annealing point

Question 8

Stringency

Answer 8

balancing sequence length for primers
- too many nt = more chance for mismatched nt
- too few = not specific enough

Question 9

PCR primer length

Answer 9

18-25 n.t. range

Question 10

average primer length

Answer 10

20 n.t.
- 2^20 different nucleotide sequences possible
- specific enough