Biochem10

Question 1

What is the technique used for amplifying a desired fragment of DNA?

Answer 1

PCR

Question 2

What are the 3 steps of PCR?

Answer 2

1-Denaturation - Heating generates 2 separate strands
2-Annealing - cooling phase, DNA primers anneal to specific sequence on each strand to be amplified
3-Elongation - DNA polymerase replicates DNA sequence following each primer

Question 3

What is used to seperate PCR products based on size?

Answer 3

Agarose gel electrophoresis

smaller molecules travel further

Question 4

Southern Blot

Answer 4

DNA

Question 5

Northern Blot

Answer 5

RNA

mRNA levels

Question 6

Western Blot

Answer 6

Protein

Question 7

Southwestern Blot

Answer 7

DNA-binding proteins (i.e. transcription factors)

Question 8

What is the mnemonic for the blotting procedures?

Answer 8

SNoW DRoP

Southern = DNA
Northern = RNA
Western = Protein

Question 9

What technique uses DNA or RNA probes to profile gene expression levels of thousands of genes simutaneously?

Answer 9

Microarrays

Question 10

What are microarrays able to detect?

Answer 10

Single Nucleotide Polymorphisms (SNPs)

Question 11

What is a rapid immunologic technique for testing antigen-antibody reactivity?

Answer 11

Enzyme-linked Immunosorbent Assay (ELISA)

Question 12

In an Indirect ELISA, the patients blood sample is probed with what?

Answer 12

Antigen

Detects if Ab is present in blood
Secondary Ab coupled to color-enzyme is added to detect first Ab

Question 13

In an Direct ELISA, the patients blood sample is probed with what?

Answer 13

Antibody

Ab is coupled with color-enzyme to see if specific Ag is present in blood

Question 14

What tecnique is used to find specific localization of genes and directly visual anomalies (ie. microdeletions) at the molecular level?

Answer 14

Fluorescence in situ hybridization (FISH)

Fluorescent DNA or RNA probe binds to specific gene site of interest on chromosomes

Question 15

What is Karyotyping?

Answer 15

Process in which METAPHASE chromosomes are stained, ordered, and numbered according to morphology, size, arm-length ratio, and banding pattern

Question 16

What technique is used to diagnose chromosomal imbalances (i.e., autosomal trisomies, sex chromosome disorders)?

Answer 16

Karyotyping

Question 17

When would you used Fluorescent in situe hybridization over Karyotyping?

Answer 17

When deletion is too small to be visualized by karyotyping