Exam 3 - Molecular Biology Experimental Methods Flashcards
Chromatography, agarose beads in the column are studded with something like an antibody or metal that selectively attracts a single antigen.
Affinity chromatography
Chromatography, proteins binds to the column under high salt conditions and are eluted under low salt condition.
Hydrophobic chromatography
Chromatography, charged proteins stick to the column, proteins are eluted by changing the pH of the buffer.
Ion exchange chromatography
Chromatography, larger proteins elute faster than smaller proteins, proteins are separated by size.
Size exclusion chromatography
Four principles to consider when making a cell culture.
Food, shleter, climate control, and companionship.
Inability to make something essential for life.
Auxotrophy
Climate control factors to consider when making a cell culture.
Temperature, humidity, pH, oxygenation, and light.
General structure of antibody.
Y-shaped, 4 proteins (i.e. 2 heavy chains and 2 light chains), 4 disulfide bonds.
Type of environment required for creation of disulfide bond.
Oxidizing environment (i.e. ER lumen and peroxisomes in eukaryotes)
Larger part of antigen that interacts with antibody.
Epitope
Lab-produced antibody with ability to recognize multiple epitopes.
Polyclonal antibody
ELISA, single Ab binds the antigen and carries the signal.
Direct ELISA
ELISA, primary Ab binds antigen, secondary Ab binds primary and carriers signal.
Indirect ELISA
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)
Sandwhich ELISA
ELISA, antigen presence is indicated by absence or decrease of signal, order of addition = Pure antigen - Primary Ab + Sample - Secondary Ab
Competition/Inhibition ELISA
Most selective column chromatography.
Affinity
Least selective column chromatography
Size exclusion
Chromatography, resin is something that specifically binds protein (i.e. antibody, metal).
Affinity
Chromatography, resin is charged.
Ion exchange
Chromatography, resin is hydrophobic.
Hydrophobic interaction
Chromatography, resin is porous agarose beads.
Size exclusion
Only blot that uses Ab
Western
Blot that detects protein
Western
Blot that detects DNA
Southern
Blot that detects RNA
Northern
Science of sequencing and the art of analysis of the genome of an individual for health related decision making.
Personal genomics
What are the following examples of - Single-nucleotide polymorphism (SNP), exome/genome sequencing?
Genetic tests
Determines if an individual has an increased risk for a particular disease.
Predicitive genetic testing
Determines if an individual has a genetic disease.
Diagnostic genetic testing
Focused on new taxonomy of human disease based on molecular biology using informatics to prevent/treat disease.
Precision medicine
Generally, how are genetic tests developed?
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.
Objective is to ensure quality genetic lab testing. Certification requires regular training/calibration/reliability and accuracy of tests.
Clinical Laboratory Improvement Amendments (CLIA)
Identifies variations in an individual’s genetic makeup to determine whether a drug is suitable for that patient.
Pharmocogenomics (i.e. Warfarin)
Significance of BRCA1 carrier.
55-65 percent chance of developing breast cancer by the age of 70.
Significance oe BRAC2 carrier.
45 percent chance of developing breast cancer by the age of 70.
When would one consider BRCA1/2 testing?
Family history - Multiple cases within the family. male breast cancer, or case in family member before 50.
Noninvasive, alteration to colonoscopy, analysis of hypemethylated of CpG islands in gene-promoter regions associated with colon cancer.
Cologuard
Ethical considerations for genetic testing.
Uncertainty of interpretations based on genotypic data alone, information can be used against individuals by insurance companies.
