MCBG 12 Flashcards
What is warfarin?
Anticoagulant NOT a blood thinner
analogue of vitamin K
What interferes with warfarin?
Vitamin K
WARFARIN. how this molecule works to slow blood clotting?
Inhibits VII
Factor VII is vitamin K dependent
Advanced molecular techniques. Analysis of DNA at the nucleotide level
DNA sequencing
Advanced molecular techniques. Analysis of DNA at the gene level
Southern hybridisation (Southern Blot + Radioactive hybrisation) Microarray PCR variations
Advanced molecular techniques. Analysis of DNA at the chromosome level
Karyotyping
FISH
What are restriction enzymes?
Molecular scissors that remove a specific sequence
Molecular techniques and diagnosis
Analysis of DNA at the gene level?
Restriction enzymes
DNA gel electrophoresis
PCR
Molecular techniques and diagnosis
Analysis of protein
Protein electrophoresis
Immunoassays
Enzyme assays
Explain Gel electrophoresis.
A sequence of Genetic material is chopped up using restriction enzymes and place in a wells in the gel (aka filter) with electrodes on either end. Turn on the current and the negatively charged pieces will move to the cathode. Small pieces will move further
How does cloning work?
Isolate sequence of GM you want. Insert it into a plasmid using a vector, recombinant DNA will produce proteins and then you isolate the recombinant DNA
Why do we clone human genes? 4
- To make useful proteins (insulin)
- Find of gene function
- Genetic screening
- Gene therapy (CF [MBP2’
What is the function of PCR?
Amplification of target DNA
How does PCR work? 3
- Denaturation - the DNA at 95oC to make it single-stranded
- Primer Annealing - the two primers bind the appropriate complementary strand
- Primer Extension - DNA polymerase extends the primer by its polymerase activity; Taq polymerase, the DNA polymerase, performed at 72oC
How does protein electrophoreisis differ from DNA.
Proteins instead of DNA
Both still Neg charged
Whats isoelectric focusing?
Proteins are seperated due to due to charge. They migrate through a pH column until they reach net neutral charge
Whats pI?
isoelectric point,” this is the pH at which a molecule has a net neutral charge
Northern blot?
Gel electrophoresis for RNA
Southern Blot?
Gel electrophoresis for DNA
Western blot
Gel electrophoresis for Proteins
Whats an Enzyme assay?
An experiment that works out ENZYMATIC ACTIVITY
How might antibodies be used?
Post western blot - used to identify specific proteins
Whats SDS gel?
Sodium dodecyl sulphate denatures proteins. Agrose and Polyacrylamide are the types of gel
Why is it important to measure enzymes?
Markers for disease
What gel do you use for proteins 5-500 bp
Polyacrylamide
What gel do you use for large DNA (50-20,000)
Agrose
