Week 9

Question 1

Molecular techniques are highly sensitive methods used to ______ genetic abnormalities and ______ disease progression

Answer 1

detect, monitor

Question 2

In RNA what base replaces T (Thymine)?

Answer 2

U (Uracil)

Question 3

What are the 4 base pairs in DNA?

Answer 3

Guanine & Cytosine

Adenine & Thymine

Question 4

3 base sequences that code for amino acids are called what?

Answer 4

Codons

Question 5

• A pairs with ____

* G pairs with _____

Answer 5

• A pairs with T

* G pairs with C

Question 6

How many different amino acids are there and how many different tRNAs

Answer 6

• 20 different amino acids

* 20 different tRNAs

Question 7

This is a tumour suppressor protein that signals damaged DNA

Answer 7

p53

Question 8

3 prime is what group and 5 prime is which group?

Answer 8

3' = OH Group
5' = Phosphate group

Question 9

DNA Synthesis always proceeds:
• Reading __’ to __’ direction
• Synthesis __’ to __’ direction

Answer 9

• Reading 3’ to 5’ direction

* Synthesis 5’ to 3’ direction

Question 10

Millions of copies of a target area will be made. Specific primers and probes are needed. This is describing what?

Answer 10

PCR - polymerase chain reaction

Question 11

In addition to positive and negative controls why is a “No DNA” control used?

Answer 11

To exclude contamination

Question 12

After 30 cycles of PCR amplification how many copies of the target sequence are created?

Answer 12

One billion
Note: A billion copies is enough to
visualise via electrophoresis

Question 13

What is more stable and easier to work with DNA or RNA?

Answer 13

DNA more stable and easier to work with than RNA

Question 14

Heat insensitive DNA polymerase from Thermus aquaticus is referred to as what ?

Answer 14

Taq polymerase

Question 15

PCR master mix contains what 2 things?

Answer 15

Taq polymerase & Nucleotides

Question 16

What are the 3 steps of the PCR amplification protocol?

Answer 16

Denaturation
Annealing (primers)
Elongation

Question 17

Briefly describe denaturaiton (include temp)

Answer 17

~95C strands separate but not destroyed

Question 18

Briefly describe annealing (include temp)

Answer 18

40 – 60C

primers(short DNA molecules) bind to flanking regions of the target DNA

Question 19

Briefly describe elongation (include temp)

Answer 19

72C
TAQ polymerase (a heat resistant bacteria) adds nucleotides to bound primers

Question 20

How many nucleotides per second added to 5’ end to create amplicon

Answer 20

1000

Question 21

Primer specificity is critical so that ______ represents area of interest

Answer 21

amplicon

Question 22

Why is a positive control in PCR used?

Answer 22

To ensure target is being amplified

Question 23

What does the negative control in a PCR lack?

Answer 23

• Lacks target sequence

Question 24

DNA has a net negative charge due to what?

Answer 24

nucleic acid phosphate groups

Question 25

What is the ladder control in gel electrophoresis?

Answer 25

Control containing different size DNA
fragments of known mass
Run alongside other controls and test
samples

Question 26

Is Standard PCR Qualitative or Quantitative

Answer 26

Qualitative

Question 27

What does Real-time PCR measure

Answer 27

Measures rate of amplification of target
Quantitative
• Viral load
• Tumour load or minimal residual disease

Question 28

What is this process called: Recognise and bind to specific nucleotide sequences, Cut the sequence at that point

Answer 28

Restriction endonucleases or restriction enzymes

Question 29

What does restriction endonucleases produce?

Answer 29

Restriction fragments

• 4 to 15 nucleotides long

Question 30

What is this process called: Technique used to determine the order in which nucleotides are
arranged in a nucleic acid sequence

Answer 30

DNA sequencing