What is a PCR array?

PCR arrays are dedicated to gene expression analysis by quantitative PCR. They are sets of primers, on the same support (384-well microplate), selected according to a theme or a given biological process. PCR arrays can also be used for the validation of results obtained after transcriptomic analysis.

How do PCR Arrays work?

The PCR array system, with high-quality input RNA, yields single bands of the predicted size without primer-dimers or other secondary products, therefore providing the highly accurate real-time PCR results (see figure ” A single gene-specific product in every reaction”).

Is RT PCR quantitative?

Quantitative RT-PCR assay is considered to be the gold standard for measuring the number of copies of specific cDNA targets in a sample but it is poorly standardized.

How do TaqMan probes work?

TaqMan probes consist of a fluorophore covalently attached to the 5′-end of the oligonucleotide probe and a quencher at the 3′-end. Degradation of the probe releases the fluorophore from it and breaks the proximity to the quencher, thus relieving the quenching effect and allowing fluorescence of the fluorophore.

How do Microarrays measure gene expression?

A microarray is a laboratory tool used to detect the expression of thousands of genes at the same time. The DNA molecules attached to each slide act as probes to detect gene expression, which is also known as the transcriptome or the set of messenger RNA (mRNA) transcripts expressed by a group of genes.

What is the basic principle of RT-PCR?

Quantitative reverse transcription PCR (RT-qPCR) is used when the starting material is RNA. In this method, RNA is first transcribed into complementary DNA (cDNA) by reverse transcriptase from total RNA or messenger RNA (mRNA). The cDNA is then used as the template for the qPCR reaction.

Is RT-PCR qualitative or quantitative?

A two-step approach using qualitative RT-PCR (for detection) and quantitative RT-PCR (for viral load quantification) is highly recommended for studies focusing on viral loads, as clearly presented by Lescure and colleagues.

What does a probe do in PCR?

Probes are fluorescently labelled DNA oligonucleotides. They are designed to bind downstream of one of the primers during the PCR reaction and to give a fluorescent signal during the reaction. The 5′ end of the probe is labelled with a fluorescent reporter molecule.

What is difference between probe and primer?

The main difference between probe and primer is that probe is that probe is used to detect the presence of a specific DNA fragment in the mixture through the hybridization with a double-stranded DNA whereas primer is used in the initiation of the polymerase chain reaction by hybridization with single-stranded DNA.