Lab Techniques I

Question 1

PCR

Answer 1

3 stages

• 1 denaturation - heating
• 2 annealing - premade DNA primers
• 3 elongation - heat stable DNA polymerase

repeated - sequence amplifies
separate using gel electrophoresis

Question 2

southern blot

Answer 2

DNA identification

Question 3

northern blot

Answer 3

RNA identification

Question 4

western blot

Answer 4

protein identification

Question 5

HIV test

Answer 5

first - ELISA

second - confirmatory - western blot

Question 6

southwestern blot

Answer 6

DNA-binding protein identifiction

Question 7

microassay

Answer 7

thousand nucleic acid sequence on grid
-DNA RNA probe hybridized

profile gene expression thousands of genes simultaneously

detect single nucleotide polymorphs and copy number variations

Question 8

ELISA

Answer 8

enzyme linked immunosorbent assay

detect specific antigen or antibody

Question 9

direct ELISA

Answer 9

test Ab for specific Ag

Ab coupled to color generating enzyme

Question 10

indirect ELISA

Answer 10

test Ag or Ab for Ab or Ag, respectively

second Ab used to bind first - this is coupled to color generating enzyme

Question 11

kayotyping

Answer 11

metaphase chromsome
-stained, ordered, numbered

for chromosomal imbalances

Question 12

FISH

Answer 12

fluocrescence in situ hybridization

fluorescent DNA or RNA probe binds specific gene site of interest

localization of gene on chromosome

Question 13

cloning

Answer 13

mRNA isolation

• reverse transcriptase - cDNA forms
• cDNA - into bacterial plasmid
• bacteria - produces cloned DNA

Question 14

cre-lox system

Answer 14

manipulate gene expression at specific development point

Question 15

RNA interference

Answer 15

dsRNA - complementary to mRNA

introduced human cells - separates and promotes degradation - target mRNA - knock down gene expression