Shotgun

Question 1

What are the steps of genomic analysis

Answer 1

Ss

Question 2

Types of most used genome sequencing methods?

Answer 2

WGS

Hierarchical shotgun method

Question 3

Explain the steps in whole-genome sequencing

Answer 3

make fragments

clone into vectors

sequence clones

assemble into contigs

assemble contigs into scaffolds

map scaffolds to chromosomes

Question 4

Differences in hierarchical shotgun sequencing compared to

Answer 4

genomic dna broken down

sub-cloned into BAC(100-200kb)

tilling path is established

those large fragments are sequenced using WGS & assembled

Question 5

What are generations in sequencing

Answer 5

1st gen - sanger

2nd gen - pyrosequencing

next gen - solexa

Question 6

Explain the steps of Sanger sequencing

Answer 6

DNA is denatured

Oligonucleotide primer is added

DNA polymerase extend the strands by adding dNTPs

this synthesis is inhibited in the presence of ddNTPs

separate rxns for 4 ddNTPs

series of extended fragments terminating at various positions

electrophorosis of four rxns

visualized on a film / capillary electrophoresis - laser excites fluorophores making color