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Flashcards in MCBG 12 Deck (26):
1

What is warfarin?

Anticoagulant NOT a blood thinner
analogue of vitamin K


2

What interferes with warfarin?

Vitamin K

3

WARFARIN. how this molecule works to slow blood clotting?

Inhibits VII
Factor VII is vitamin K dependent

4

Advanced molecular techniques. Analysis of DNA at the nucleotide level

DNA sequencing

5

Advanced molecular techniques. Analysis of DNA at the gene level

Southern hybridisation (Southern Blot + Radioactive hybrisation)
Microarray
PCR variations

6

Advanced molecular techniques. Analysis of DNA at the chromosome level

Karyotyping
FISH

7

What are restriction enzymes?

Molecular scissors that remove a specific sequence

8

Molecular techniques and diagnosis
Analysis of DNA at the gene level?

Restriction enzymes
DNA gel electrophoresis
PCR

9

Molecular techniques and diagnosis
Analysis of protein

Protein electrophoresis
Immunoassays
Enzyme assays

10

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

11

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

12

Why do we clone human genes? 4

1. To make useful proteins (insulin)
2. Find of gene function
3. Genetic screening
4. Gene therapy (CF [MBP2'

13

What is the function of PCR?

Amplification of target DNA

14

How does PCR work? 3

1.Denaturation - the DNA at 95oC to make it single-stranded

2. Primer Annealing - the two primers bind the appropriate complementary strand

3. Primer Extension - DNA polymerase extends the primer by its polymerase activity; Taq polymerase, the DNA polymerase, performed at 72oC

15

How does protein electrophoreisis differ from DNA.

Proteins instead of DNA
Both still Neg charged

16

Whats isoelectric focusing?

Proteins are seperated due to due to charge. They migrate through a pH column until they reach net neutral charge

17

Whats pI?

isoelectric point,” this is the pH at which a molecule has a net neutral charge

18

Northern blot?

Gel electrophoresis for RNA

19

Southern Blot?

Gel electrophoresis for DNA

20

Western blot

Gel electrophoresis for Proteins

21

Whats an Enzyme assay?

An experiment that works out ENZYMATIC ACTIVITY

22

How might antibodies be used?

Post western blot - used to identify specific proteins

23

Whats SDS gel?

Sodium dodecyl sulphate denatures proteins. Agrose and Polyacrylamide are the types of gel

24

Why is it important to measure enzymes?

Markers for disease

25

What gel do you use for proteins 5-500 bp

Polyacrylamide

26

What gel do you use for large DNA (50-20,000)

Agrose