module 6 manipulating genomes

Question 1

outline the production of a DNA profile

Answer 1

extract DNA and multiply using PCR
cut strands with restriction endonuclease
separate using gel electrophoresis
add fluorescent probes (hybridisation)
use UV light to see tags

Question 2

outline steps of PCR

Answer 2

95C for 30 seconds to break H bonds
55c add primers to bind to ends
75c for one minute for DNA polymerase to add bases and build complementary strands

Question 3

outline DNA sequencing

Answer 3

add primers, DNA polymerase, and nucleotides
heat to 95c to break h bonds and
50c to allow primers to bind to strands
60C so DNA polymerase can build strands
add terminators bases that result in different length strands
split by length using capillary sequencing fed into a computer to anaylse

Question 4

isolating desired genes (insulin)

Answer 4

mRNA extracted from pancreatic cells
treated with reverser transcriptase (cDNA)
the plasmid obtained from bacteria and cut with restrictions enzymes
DNA ligase fuse plasmid and cDNA
recombinant plasmid introduced to host cells then leave to multiply

Question 5

somatic gene line therapy

Answer 5

replacing mutant alleles with healthy alleles

Question 6

germline therapy

Answer 6

insert healthy allele into an egg cell