Real Time PCR Vs Traditional PCR: A Guide to Choosing the Right One for Your Lab

 

Polymerase chain reaction or PCR is a technique used in molecular biology to analyse and enlarge RNA and DNA samples. It is widely used for conducting research, maintaining quality control, and in the field of diagnostics. However, when buying a PCR machine, you may feel confused when choosing between traditional or real-time machine. There are differences between traditional or endpoint PCR and real-time or quantitative PCR machine. You need to know the differences between the devices to understand which one is the right fit for your laboratory.

What is a traditional PCR machine?

Traditional PCR is a well-established and familiar technique that has been used in laboratories for decades. It helps augment the RNA and DNA sequence through repetition of cooling and heating cycles. That way, the DNA strands become separated and aid in DNA replication. Once the cycles are completed, the visualisation process takes place through gel electrophoresis.

The products that have been amplified are then stained with a dye; thereafter the intensity of bands on the gel shows the quantity of enlarged DNA. Although the traditional PCR machine performs reliably and has been used in research, forensic science, and diagnostics for a long time, it fails to meet the gaps when it comes to quantitative analysis and real-time data acquisition. So, when buying the best PCR machine for labs, you need to analyse the features and specifications of both to make an informed decision.

What is a real-time PCR?

A real-time PCR machine also called quantitative PCR scores over the traditional machine in several different ways. When using this machine, you need not wait until the completion of PCR to analyse and amplify DNA but come to know about the quantification when the DNA is amplified. This is possible through the monitoring of fluorescence that emits during the amplification process and provides continuous data in real-time. The fluorescence generates due to specific dyes that usually bind to the target DNA. Now, when the reaction progresses further, the fluorescence emitted binds with the quantity of DNA that undergoes amplification and eliminate the need to wait for the completion of PCR.

Undoubtedly, real-time PCR is a highly versatile method and extensively used for gene expression analysis, detection of mutation, viral load, and more.  The ability of this machine to measure DNA and RNA in real-time makes it a valuable tool for clinicians and researchers.

Understanding the key differences between traditional and real-time PCR

Here are the differences between real-time and traditional PCR machine:

• Quantification

1. The traditional PCR is limited when providing qualitative data and helps you identify whether a particular DNA sequence is present or absent in the sample. However, it cannot tell the amount of DNA in real-time and you need to wait till the completion of PCR analysis. Naturally, the quantification of DNA is not precise.
2. Real time PCR helps in precise quantification of data. The fluorescence that emits during the process of amplification allows researchers measure the amount of RNA and DNA within the samples at each cycle.

 

• Detection method

1. The traditional PCR is a highly sensitive machine and can detect even the lowest amount of DNA but it is restrained by post-PCR analysis. Naturally, it cannot detect low quantity of the target DNA.
2. Real-time PCR is relatively more sensitive compared to the traditional PCR. As DNA amplification is constantly monitored, DNA and RNA can be detected early in low-quantity samples.

 

• Effective timing

1. Traditional PCR takes a few hours to complete and you need to wait until post-PCR analysis to visualise the results. Naturally, you need more time to complete the entire experiment.
2. Real-time PCR delivers quick results as the quantification and amplification process takes place simultaneously. So, there is no need for you to wait until post PCR analysis. Naturally, the time taken is less, especially for high-throughput applications.

 

• Cost

1. The traditional PCR machines are more affordable compared to real-time PCR systems. However, you need to spend additionally for consumables, gel electrophoresis reagents like agarose gel and DNA staining agents. Therefore, the traditional PCR machines may add to the cost due to these requirements.
2. Real-time PCR machines are more expensive than traditional machines in terms of investment and maintenance but the accuracy of data justifies the cost to a great extent. So, if you analyse the real-time PCR machine benefits that way, making an informed decision is easier.

When choosing between traditional and real-time PCR machines, a lot depends on the specific requirements of the laboratory. So, if you are working with samples with low RNA and DNA abundance, real-time PCR machine is the right choice. However, if you are conducting basic DNA analysis where the most important thing is the absence and presence of a specific DNA sequence, the traditional PCR machine is more important. At IGene Labserve, we sell top-notch quality of laboratory equipment. Just explore the models here https://www.igenels.com/ and order the equipment you need right away.