Chapter 10

Question 1

polyclonal vs monoclonal antibodies

Answer 1

poly- antibodies bind to different epitopes on the same protein

mono- homogeneous antibodies bind only to a single epitope (one specific antigen)

Question 2

PCR

Answer 2

used to amplify DNA
-increases the amount of DNA so is useful when there’s only trace amounts available
- very efficient and specific

• need specific design primers
  -heat resistant DNA polymerases allows to perform PCR

Question 3

qPCR

Answer 3

real-time quantitative polymerase chain reaction:
- amplify AND quantify a DNA sequence
-measures quantity of DNA produced by each PCR cycle
- fluorescents bind to new DNA copies as they’re made

Question 4

reverse transcription PCR (RT-PCR)

Answer 4

PCR for RNA
-RNA into cDNA then cDNA goes thru PCR

Question 5

blotting

Answer 5

-identify target fragments

Southern - DNA (fluorescent probes)
Northern - RNA (fluorescent probes)
Western - Proteins (antibodies)

Question 6

2D gel electrophoresis

Answer 6

seperate proteins based on size and isoelectric point

Question 7

GFP fusion protein

Answer 7

protein attached to a green fluorescent protein / both expressed in a cell allowing for visualization

Question 8

RNA seq.

Answer 8

quantifies gene expression by quantifying RNA transcripts

-RNA->cDNA->cDNA then sequenced using next gen methods

*don’t need to choose probes ahead of time / examine non-coding RNAs

Question 9

Microarrays

Answer 9

compares mRNA expression between two samples
-mRNA hydrbidizes to cDNAs that are bound to spots on glass slides
-bias

Question 10

what is a key difference between microarray and RNA-seq

Answer 10

you must select genes of interest for micrarrays but NOT for RNA-seq

Question 11

knockout mice symbols

Answer 11

+/+ wild type
-/+ one allele knocked out
-/- both alleles knocked out

Question 12

which techniques can be used for gene expression (RNAs)

Answer 12

-RNA seq
-Microarrays
-Gel electro and northern blotting
-RTqPCR

Question 13

in situ hybridization

Answer 13

allows a specific nucleotide sequence (DNA or RNA) to be visualized in its normal cellular location (where it’s being expressed)

time and place mRNA product of a gene is being expressed

• uses probes

Question 14

CRISPR/Cas9

Answer 14

edits parts of genome by removing/adding or altering sections of the DNA sequence

-uses a specifically designed RNA molecule to guide an enzyme called Cas9 to a specific sequence of DNA

Cas9= endonuclease which causes double stranded DNA to break allowing it to then be modified

-create animal models of disease

Question 15

RTqPCR

Answer 15

amplify and quantify cDNA