Next Gen Sequencing

Question 1

What is next gen sequencing ?

Answer 1

Uses human genome project as a reference and just checks variable region

Question 2

emulsion PCR

Answer 2

emPCR is done in an oil-water emulsion to create an aqueous bubble around each bead

Question 3

Pyrosequencing

Answer 3

luciferase is added to the picotitre plate in addition to sulphrylase
wash over with different base and CCD will capture flashes when it adds the base

Question 4

sanger sequencing (not next gen)

Answer 4

replicate DNA with some chain terminating nucleotides
repeat separately with each nucleotide
southern blot and read

Question 5

shotgun sequencing (not next gen)

Answer 5

fragment and squence.

use computer and find overlap

Question 6

cellularity

Answer 6

the amount of cells in a tumour that are normal

Question 7

using next gen to analyse metastasis

Answer 7

sequence genes of tumour and metastasized tumour.

one showed deleted gene; indicative of loss of function

Question 8

why are lymphomas common ?

Answer 8

the hypervariable region of antibodies require a lot of mixing and crossing over. this means that cancer genes are easily formed

Question 9

breakapart assay

Answer 9

do in situ hybridization (FISH)
put in a fluorescent probe, one end red, one end green
If they break apart (cross over, loss of one side etc) will not appear yellow