Name the individual who developed the polymerase chain reaction.
List the 5 components required for the polymerase chain reaction and
the function of each component.
Briefly describe the 3 steps in the polymerase chain reaction (PCR).
State the optimal annealing temperature.
State the optimal temperature of replication/primer extension.
State the difference between the two-step PCR and three-step PCR.
State the optimal length of primers.
State the binding direction of the forward and reverse primers.
Describe the problem of primer-dimer formation.
List 6 potential sources of DNA template in a clinical sample.
State the amount of DNA required for a successful PCR reaction.
Describe the conditions which must be met for the best DNA template.
State the source of Taq polymerase and describe the advantage of using
it in a PCR procedure.
List the components of the PCR buffer and the function of each.
Describe a thermal cycler's function and the advantage this has over
manual methods.
State the importance of having a separate area for sample preparation,
pre-PCR work, post-PCR work and analysis of results..
State how the PCR product is analyzed.
Given a PCR agarose gel identify: misprime, PCR product and primer
dimer bands.
List and describe the purpose of the three controls which are utilized to ensure the accuracy of the PCR run.
List and describe 4 physical and chemical methods used to control PCR contamination and
state why this is so important.
Define "mispriming" and breifly describe 3 hot-start set
ups which may prevent or decrease this from occurring.
BRIEFLY describe the following PCR modification procedures: Multiplex
PCR, Reverse Transcriptase PCR, nested PCR, real time PCR and arbitrarily
primed PCR.
Briefly describe the principle of transcription-based amplification systems.
Briefly describe the following probe amplification techniques: ligase
chain reaction strand displacement amplification and QB replicase.
Briefly describe the following signal amplification methods: branched
DNA amplification, hbrid capture assays, cleavage based amplification
and cycling probe.
