PCR Flashcards
How is qPCR different from PCR?
In qPCR vs PCR, the key differences are quantification, speed and resolution.
End point PCR enables qualitative or semi-quantitative analysis at the end of all PCR cycles via an agarose gel or microchip.
qPCR relies on fluorescent dyes or probes and calibration curves to deliver quantitative data in real-time.
What is the purpose of a qPCR?
In research laboratories, qPCR assays are widely used for the quantitative measurement of gene copy number (gene dosage) in transformed cell lines or the presence of mutant genes.
What is the process of PCR?
The polymerase chain reaction is a nucleic acid amplification testing procedure
Consists of denaturing, renaturing, elongating, and amplifying a short segment of DNA or RNA.
Implemented by incorporating DNA I polymerase, which is derived from Thermus aquaticus, also known as Taq polymerase.
What are Primers?
-Short, specific DNA sequences.
-are like starting points for making copies of a specific DNA segment.
How long are primers?
18-25 nucleotides long
PCR uses 2 primers:
-Forward Primer: binds to the upstream (5’) region of the target sequence
- Reverse Primer: binds to the downstream (3’) region of the target sequence
PCR components
-DNA sample
-Primers
-Nucleotides
-Taq polymerase
-Mix buffer
-PCR tube
QF-PCR stands for
Quantitative Fluorescent Polymerase Chain Reaction
QF-PCR is used for?
It’s a technique used in molecular biology to rapidly diagnose chromosomal abnormalities, particularly in prenatal testing.
While automated thermocyclers are the dominant method for PCR today, there are a few reasons why someone might still choose to perform manual PCR:
-Limited Resources
-Educational Purposes
-Specific Experimental Needs
-Equipment Failure
An amplicon
is a piece of DNA or RNA that is the source and/or product of natural or artificial amplification or replication events.
In molecular biology, the term typically refers to the product of a polymerase chain reaction (PCR) amplification process.
