Principe of traditional PCR is applied, but PCR products are detected at each cycle in real time as reaction progresses

Quantitation of nucleic acid must be done at the log-linear phase

Amount of target nucleic acid is reported as a copy number of concentration

Calculated from a standard curve as generated for absolute quantification

This was discovered in reverse transcribing viruses

This assay combine reverse transcription and PCR in a single tube and buffer, using a reverse transcriptase along with a DNA polymerase

RNA- dependent DNA polymerases which bind to RNA templates and convert them to their complementary DNA sequence in the presence of dNTPs and other essential cofactors.

Real time PCR is also known as?

How many major phases are there in the PCR amplification curve?

Exponential amplification becomes detectable

Basal level of fluorescence defined during initial stage of PCR

Threshold cyle, this is the number at which the fluorescence signal crosses the threshold

Point which significant and specific amplification occurs and fluorescence signal rises above backround level

This assay the reverse trancription and PCR steps are performed in separate tubes, with different optimized buffers, reaction conditions, and priming strategies

In threshold an C(t), the more initial concentration, the less the C(t) value

Reactions slows down due to depletion of reagent components and lower PCR efficiency

PCR is used to amplify DNA, while RT-PCR is used to amplify RNA

Amount of target nucleic acid is reported comparative to the abundance of another gene

This is used in a variety of applications including gene expression analysis, RNAi validation, microarray validation, pathogen detection, genetic testing, and disease research

What are the 4 major phases in the PCR amplification curve?

This is used to detect and quantify RNA. Total RNA or mRNA is first transcribed into complementary DNA (cDNA).

Reaction components become limited; reaction stops

Start of PCR, amplification is not detectable.