PCR

Question 1

Uses of PCR

Answer 1

DNA fingerprinting, classification of organisms, genotyping, clinical diagnostics, detections of pathogens, genetic cloning,

Question 2

PCR primer sequences

Answer 2

should always be found on opposite strands, and oriented towards each other

Question 3

DNA primers anneal to

Answer 3

complementary strands

Question 4

variations in the size of an unstable allele in huntingtons disease.

Answer 4

causes variable expressivity.

Question 5

PCR can be used to amplify short pieces of DNA

Answer 5

AND size differences can be visualised by gel electrophoresis.

Question 6

DNA amplification by molecular cloning

Answer 6

steps are:

1. DNA fragmentation, small enough to fit in vector
2. Ligation in vector, plasmids or YAC
3. Transformation: introduction of vector in bacteria or yeast
4. culture and screen: antibiotic resistance

Question 7

Recombinant plasmids are amplified in and purified

Answer 7

from a host cell

Question 8

Recombinant plasmids are amplified in and purified

Answer 8

from a host cell

Question 9

Cloning vector requirements

Answer 9

convenient restriction sites
Autonomous replication
selectable marker
Some plasmid vectors are manipulated to permit screening for recombinants (lacz for blue white screening)

Question 10

Genomic libraries contain clones, which, together

Answer 10

represent the entire genome

Question 11

Genomic DNA is cut with

Answer 11

a restriction enzyme

Question 12

cDNA libraries represent

Answer 12

genes which are expressed

Question 13

Colony hybridisation allows

Answer 13

purification of the clone of interest

Question 14

The ideal genome editing tools would be

Answer 14

Edit any genome tools, high efficiency, high DNA sequence specificity, no undesired by- products

Question 15

Cas9 is an

Answer 15

RNA guided DNA endonuclease: Cas9 endonuclease forms a complex with a guide RNA and localises to a target DNA sequence following a gRNA: genomic DNA pairing.

Question 16

Restriction fragment length polymorphisms can

Answer 16

distinguish between 2 alleles (hetero and homo zygous)

Question 17

Restriction fragment length polymorphisms are

Answer 17

abundant throughout eukaryotic genomes. RFLP’s result from differing positions of restriction sites

Question 18

The bands on the gel when RNA is run represents

Answer 18

the small and big ribosomal RNA subunit

Question 19

Probes from a variety of sources will

Answer 19

hybridise if the sequence similarity is high

• polymorphic alleles
• related genes or sequences from same species
• same sequence different species

Question 20

Northern analysis reveals

Answer 20

gene expression data

Question 21

RNA sequences can be used for

Answer 21

alternative gene spliced transcripts, post translational modifications, differences in gene expression in the whole transcriptome,