Search results
(A) RT-PCR workflow. RNA is isolated and cDNA is generated via reverse transcription (RT); PCR is then carried out to amplify areas of interest. (B) qPCR schematic. DNA is isolated and...
- Zaloguj sie
(A) RT-PCR workflow. RNA is isolated and cDNA is generated...
- Help Center
© 2008-2024 ResearchGate GmbH. All rights reserved. Terms;...
- Zaloguj sie
Quantitative reverse transcription polymerase chain reaction, also called RT-qPCR, is used to detect and quantify RNA. Total RNA or mRNA is first transcribed into complementary DNA (cDNA). The cDNA is then used as the template for the quantitative PCR or real-time PCR reaction (qPCR).
1.1 What qPCR is & how it works Quantitative PCR (qPCR) is one of the foremost methods for detection and quantification of nucleic acid targets. The technique is also known as real-time qPCR and is not to be confused with RT-qPCR (Reverse Transcription-qPCR). It relies on the same basic reagents, enzyme and thermocycling condi-
The reverse transcriptase (RT) is as critical to the success of qRT-PCR as the DNA polymerase. It is important to choose an RT that not only provides high yields of full-length cDNA but also has good activity at high temperatures. High-temperature performance is also very important for tackling RNA with
The quantitative Reverse Transcription Polymerase Chain Reaction, abbreviated as qRT PCR is a major advancement of the Polymerase Chain Reaction (PCR) technology. Of all the methods available, qRT PCR is the most rapid, sensitive and accurate methodology for measurement of RNA.
1.1 Key Concepts of Real-Time PCR 1.1.1 What Is Real-Time PCR? In conventional PCR, the amplified product, or amplicon, is detected by an end-point analysis, by running DNA on an agarose gel after the reaction has finished. In contrast, real-time PCR allows the accumulation of amplified product to
The basic formula applying to qRT-PCR aims to convert the number of cycles at a threshold level of fluorescence (more generally termed the quantification cycle or Cq; Hellemans et al., 2007) into a relative quantity of input template present at the start of the PCR.
