Real Time PCR: Quantitative RT PCR

2007-07-18 16:23:30

( Education )



The polymerase chain reaction (PCR) is a laboratory method that allows scientists to make multiple copies of a short strand of DNA or RNA. This is useful in molecular biology laboratories, where scientists require large amounts of DNA or RNA to effectively carry out their experiments.

Excessive amounts of DNA or RNA, however, can also affect the outcome of experiments. To measure the amount of DNA or RNA in a PCR reaction, scientists carry out a real time polymerase chain reaction, or real time PCR. Specifically, quantitative RT PCR allows scientists to monitor the increase in the amount of DNA or RNA in the PCR reaction mixture. Quantitative RT PCR has many applications in various scientific fields.

Real Time PCR

Quantitative RT PCR is carried on in a real time PCR machine. The real time PCR protocol differs from ordinary PCR protocol in the structure of its free nucleotides and primers. In conventional PCR, free nucleotides and primers are not labeled with any chemicals or fluorophores. In quantitative RT PCR, however, free nucleotides and primers are attached, or conjugated to fluorescent molecules.

These fluorescent molecules are detected by a beam of light emitted by the real time PCR machine. The stronger the fluorescence emitted, the more DNA or RNA molecules there are.

Quantitative RT PCR

Quantitative RT PCR is used for various purposes in the laboratory. It can be used to measure the original amount of DNA or RNA in a sample. This is useful when evaluating the efficiency of DNA or RNA extraction protocols. This is also useful in quantifying the activity of a gene. That is, the greater the amount of RNA in a sample, the higher stronger the fluorescence emitted, and the higher the level of gene expression.

Quantitative RT PCR is also used to automatically calculate the final concentration of DNA or RNA in a sample. Because the accuracy of subsequent experiments depends on the concentration of DNA or RNA in a sample, scientists use quantitative RT PCR to know how they need to dilute the sample in order to achieve a desirable final concentration.

If you are interested in quantitative RT PCR for your research work, study its protocols, which are available online. Compare prices among different machines and packages, and choose a real time PCR machine that will fit your budget and needs. Quantitative RT PCR can be valuable to laboratories that require precision techniques to make their experiments work.


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