Exam 3 - Molecular Biology Experimental Methods

Question 1

Chromatography, agarose beads in the column are studded with something like an antibody or metal that selectively attracts a single antigen.

Answer 1

Affinity chromatography

Question 2

Chromatography, proteins binds to the column under high salt conditions and are eluted under low salt condition.

Answer 2

Hydrophobic chromatography

Question 3

Chromatography, charged proteins stick to the column, proteins are eluted by changing the pH of the buffer.

Answer 3

Ion exchange chromatography

Question 4

Chromatography, larger proteins elute faster than smaller proteins, proteins are separated by size.

Answer 4

Size exclusion chromatography

Question 5

Four principles to consider when making a cell culture.

Answer 5

Food, shleter, climate control, and companionship.

Question 6

Inability to make something essential for life.

Answer 6

Auxotrophy

Question 7

Climate control factors to consider when making a cell culture.

Answer 7

Temperature, humidity, pH, oxygenation, and light.

Question 8

General structure of antibody.

Answer 8

Y-shaped, 4 proteins (i.e. 2 heavy chains and 2 light chains), 4 disulfide bonds.

Question 9

Type of environment required for creation of disulfide bond.

Answer 9

Oxidizing environment (i.e. ER lumen and peroxisomes in eukaryotes)

Question 10

Larger part of antigen that interacts with antibody.

Answer 10

Epitope

Question 11

Lab-produced antibody with ability to recognize multiple epitopes.

Answer 11

Polyclonal antibody

Question 12

ELISA, single Ab binds the antigen and carries the signal.

Answer 12

Direct ELISA

Question 13

ELISA, primary Ab binds antigen, secondary Ab binds primary and carriers signal.

Answer 13

Indirect ELISA

Question 14

ELISA, both Ab bind antigen at different epitopes, capture Ab is absorbed to the well, primary Ab binds to antigen and carries the signal. (i.e. order goes capture Ab - antigen - primary Ab)

Answer 14

Sandwhich ELISA

Question 15

ELISA, antigen presence is indicated by absence or decrease of signal, order of addition = Pure antigen - Primary Ab + Sample - Secondary Ab

Answer 15

Competition/Inhibition ELISA

Question 16

Most selective column chromatography.

Answer 16

Affinity

Question 17

Least selective column chromatography

Answer 17

Size exclusion

Question 18

Chromatography, resin is something that specifically binds protein (i.e. antibody, metal).

Answer 18

Affinity

Question 19

Chromatography, resin is charged.

Answer 19

Ion exchange

Question 20

Chromatography, resin is hydrophobic.

Answer 20

Hydrophobic interaction

Question 21

Chromatography, resin is porous agarose beads.

Answer 21

Size exclusion

Question 22

Only blot that uses Ab

Answer 22

Western

Question 23

Blot that detects protein

Answer 23

Western

Question 24

Blot that detects DNA

Answer 24

Southern

Question 25

Blot that detects RNA

Answer 25

Northern

Question 26

Science of sequencing and the art of analysis of the genome of an individual for health related decision making.

Answer 26

Personal genomics

Question 27

What are the following examples of - Single-nucleotide polymorphism (SNP), exome/genome sequencing?

Answer 27

Genetic tests

Question 28

Determines if an individual has an increased risk for a particular disease.

Answer 28

Predicitive genetic testing

Question 29

Determines if an individual has a genetic disease.

Answer 29

Diagnostic genetic testing

Question 30

Focused on new taxonomy of human disease based on molecular biology using informatics to prevent/treat disease.

Answer 30

Precision medicine

Question 31

Generally, how are genetic tests developed?

Answer 31

A small study group identifies a genetic variation associated with disease, a larger study is done to increase sensitivity and specificty of the test to meet FDA requirements.

Question 32

Objective is to ensure quality genetic lab testing. Certification requires regular training/calibration/reliability and accuracy of tests.

Answer 32

Clinical Laboratory Improvement Amendments (CLIA)

Question 33

Identifies variations in an individual's genetic makeup to determine whether a drug is suitable for that patient.

Answer 33

Pharmocogenomics (i.e. Warfarin)

Question 34

Significance of BRCA1 carrier.

Answer 34

55-65 percent chance of developing breast cancer by the age of 70.

Question 35

Significance oe BRAC2 carrier.

Answer 35

45 percent chance of developing breast cancer by the age of 70.

Question 36

When would one consider BRCA1/2 testing?

Answer 36

Family history - Multiple cases within the family. male breast cancer, or case in family member before 50.

Question 37

Noninvasive, alteration to colonoscopy, analysis of hypemethylated of CpG islands in gene-promoter regions associated with colon cancer.

Answer 37

Cologuard

Question 38

Ethical considerations for genetic testing.

Answer 38

Uncertainty of interpretations based on genotypic data alone, information can be used against individuals by insurance companies.