Dr. Singh is currently working as a staff scientist and Head of the Infectious Diseases Laboratory (IDL) at National Institute of Immunology (NII), New Delhi. He has worked as scientist at International Centre for Genetic Engineering and Biotechnology, New Delhi and New York University Medical Centre, before coming to NII.

With the increase in demand of these PCR thermocyclers from the researchers for shorter and more accurate PCR protocols have resulted in beneficial changes to the basic thermocycler model.

Current advancements

The constantly growing competition among thermal cycler brands has given enough reasons to be happy. These new developments focus on using lower volumes of resources to give maximum yield. Initially, the lower sample volume had a risk of evaporation that negatively affected results by skewing up the concentrations of various components of a PCR reaction thereby adversely affecting the amplification of desired target. To overcome this problem, the new PCR machines came up with a new lid made up of malleable material that moulds up during the reaction, forming, a tight seal, thereby reducing evaporation.

Landmark innovation- The real time PCR

The traditional PCR machines usually measure the yield at the end point. Since the whole reaction works as a curve starting from the lag phase to exponential and then to a plateau, finally leading to a decline phase, it was always difficult to quantify the accurate product yield relative to the reactants and the reagents added during the start of the PCR.

For the sake of accuracy and precision, it is necessary to collect quantitative data at a point in which every sample is in the exponential phase of amplification (since, it is only in this phase that amplification is extremely reproducible). But, this whole process becomes really cumbersome. Real-time PCR automates this otherwise laborious process by quantifying reaction products for each sample in every cycle. The result is an amazingly broad 107-fold dynamic range, with no user intervention or replicates required. While the end point measurement of traditional PCR through agarose gel is a poor ten-fold, real time thermal cyclers can detect as little as two-fold change along with data analysis, including standard curve generation and copy number calculation, which is performed automatically.

Applications of real time PCR

Real time PCR has become a benchmark in the research field to provide quantitative measurements of gene transcription. The technology may be used in determining how the expression of a particular gene changes over time in response to a pharmacological or a pathogenic agent or change in other conditions. mRNA expression levels which are very small, need high sensitivity for measurement that is provided by this instrument. The real time PCR has received widespread positive opinions from the research field and has already become one of the basic instruments while setting up a research laboratory. This has ensured that this technique is here to stay and will go from strength to strength with constant upgrades and modifications until some new radical innovation comes out in the market.