Genetic lab tests

Question 1

PCR steps

Answer 1

Denature w/ heat
Anneal – cool w/ DNA primers, site for DNA polymerase
Elongation – amplify
Gell electrophoresis

Question 2

Southern Blot

Answer 2

“down south”
DNA sample + DNA probe
Fluorescence or XR to visualize probe

Question 3

Northern Blot

Answer 3

“Rude Yankees”

RNA sample + DNA probe

Question 4

Western Blot

Answer 4

“Beef”

Protein sample + Ab probe

Question 5

Southwestern blot

Answer 5

DNA binding protein sample + oligonucleotide probe

Question 6

Gene expression tests

Answer 6

Western blot for protein

Northern blot for RNA sequence

Question 7

Direct vs Indirect ELISA

Answer 7

Direct: tests for Ag w/ known Ab

Ag: blood sample binds if Ag present in blood
Ab on well surface that binds the Ag in blood
Anti-Ag Ab added after blood – if Ag present, causes color change

Indirect: test Ab to the Ag – known Ag added
-Ag in well, pt’s Ab bind, Ab against pt Ab’s added color change if Ab present

ex: HIV ag used to test HIV Ab – confirm with western blot

Question 8

FISH

Answer 8

Fluorescent DNA or RNA probe
Bind gene of interest
Glow if present, absent = deletion or not present

Question 9

Cloning

Answer 9

mRNA –reverse transcriptase -> cDNA
cDNA into bacterial plasmid

bacteria propagates -> multiple copies of cDNA

Coding portion of gene (cDNA) not identical to DNA

Question 10

Modifying gene expression

Answer 10

Experimental animal

Knock out: delete gene from genome
Knock in: insert gene into genome

Constitutive insertion – insert gene randomly
Conditional insertion – targeted insertion

Question 11

Karyotyping

Answer 11

Metaphase Chr sorted by paris
Chromosomal imbalances
Can do inutero