Molecular Techniques

Question 0

What is the underlying principle of molecular techniques?

Answer 0

Use the nucleus acid of the test organism as a target for analysis
DNA- codes for organism
RNA- codes for protein

Question 1

When are molecular techniques used?

Answer 1

Identification of species
Sub typing within a species
Identification of virulence factors
Analysis of antibiotic resistance genes

Question 2

What is the purpose of molecular analysis?

Answer 2

Determine whether an organism possesses the target sequence you are asking for (possession of a gene)

Question 3

What is PCR ?

Answer 3

Polymerase chain reaction
Diagnose genetics defects, detect aids virus in human cells
Also used by criminologists
Evolutionary studies

Question 4

What do we do to perform molecular analysis to determine whether the organism possesses virulence factors or antibiotic resistance genes?

Answer 4

DNA extraction - proteinase
RNA extraction - DNAse
Presence of specific gene (PCR)
Eppendorf - template DNA, free nucleotides, Taq enzyme, reaction buffer (MgCl- provide ionic stabilisation for reaction)
Gel electrophoresis - DNA conc and purity

Question 5

What is the mechanism of PCR?

Answer 5

Template 
Primers 
DNA double helix denatures 
Temp lowered primers bind to complementary portions of DNA sample 
Taq- add nucleotides 
Appropriate sequence 
(denaturation, annealing,extension, amplification, finish/hold )
Exponential amplification

Question 6

What is the name of the machine that conducts PCR?

Answer 6

Thermocycler