Diagnostic Virology

Question 1

4 steps of Western-blot method?

Answer 1

Method used to assess if patients have antibodies specific to HTLV-1 proteins 1. Separation- smaller proteins migrate faster 2. Transfer- protein bands created 3. Staining- Membrane incubated with human serum (AB), wash and then add enzymes 4. Visualisation- colour, precipitation or fluorescence reaction

Question 2

Positive results example of Western-blot

Answer 2

Question 3

3 mains tages of PCR?

Answer 3

Denaturing dsDNA at 95 C

Primer annealing at 55 C

Extending DNA strands at 72 C

Question 4

What 5 key components must be included in a PCR mix?

Answer 4

DNA template

Primers pair

TAQ polymerase

Reaction buffer

Deoxynucleotides

Question 5

How do you estimate the size of your PCR product on the agarose gel?

Answer 5

Run a DNA ladder alongside your PCR reactions

Question 6

What is the purpose of adding a loading buffer with dye to the PCR sample before loading on an agarose gel?

Answer 6

Visualise which wells contain samples

Visualise how far the gel has migrated

Makes samples sink into wells