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Comparison of PCR, RT-PCR&Real-time Fluorescent Quantitative PCR
2020-12-25 14:45
PCR amplification instrument, also known as PCR gene amplification instrument, PCR nucleic acid amplification instrument, polymerase chain reaction nucleic acid amplification instrument, is a kind of instrument that uses PCR (polymerase chain reaction) technology to amplify specific DNA , which is widely used in medical and biological laboratories.
PCR (polymerase chain reaction) has the largest category among these three technologies and is the most basic technology, which use DNA polymerase and DNA oligomer primers to replicate and amplify the DNA template. Other PCR technologies are derivative applications of PCR.
RT-PCR (reverse transcription PCR) is is aimed at RNA, mainly the application of mRNA.. The template uses DNA recorded by RNA reversal, so the information obtained is about RNA. It is generally believed that ordinary RT-PCR is semi-quantitative, that is, it can reflect a certain dose level, but it cannot be completely quantified. There are many reasons why it cannot be fully quantified, one of which is the number of cycles. After a large number of cycles, the product amplification deviates from the template dose and becomes uncontrollable. To solve this problem, the technology for quantitative analysis is real-time PCR.
Real-time PCR, fluorescence refers to the generally used detection method, quantitative is a description, generally called real-time PCR, sometimes also called quantitative PCR (quantitative PCR). The principle is that in the early cycles of amplification, the product dose and template dose have a good linear relationship, so the amount of product can be used to describe the amount of template. But the problem with this technology is that the absolute dose of the template cannot be directly measured, and the amount can only be described by comparison. In addition, although it is often used to detect RNA dose, real-time PCR and RT-PCR are not necessarily related. There are many applications for quantitative detection of specific DNA content.
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