Lecture 49

Question 1

What is FISH?

Answer 1

fluorescence in Situ Hybridization

Question 2

What is the purpose of FIHS?

Answer 2

identify genes or chromosomes in metaphase or interphase cells

Question 3

What are the steps to FISH?

Answer 3

1) collect cells (blood, embryo, etc)
2) spread chromosomes on slide
3) add DAPI (makes DNA glow blue)
4) add fluorescent probes made from BAC clones (each BAC is one coloured signal, and is 100 000 bp)

Question 4

How is a hybridization probe made?

Answer 4

BAC + labelled NT = hybridization probe

Question 5

Purpose of FISH with centromere probes?

Answer 5

count chromosomes

Each probe = 1 BAC, 1 colour

Question 6

Purpose of FISH with gene probes?

Answer 6

determine location and presence of a gene or region

paint each end of a chromosome

Question 7

Purpose of FISH with chromosome paint probes?

Answer 7

identify chromosome rearrangements

made from many BACs covering an entire chromosome arm

Question 8

What is multicolour FISH?

Answer 8

chromosomes paint each chromosome