PCR might have been the big discovery of 1983, and now a researcher has developed a new technique, a step ahead of it. Shalini Gupta reports
In 1983, Dr Kary Mullis developed a breakthrough technology, PCR, that helped scientists to better understand the functions of a living cell by amplifying the DNA many times over. The invention went on to get Mullis a Nobel Prize. PCR findings confirm that traditional culture-based analyses miss many bacteria that fail to grow readily. The sensitivity provided by these nucleic acid amplification technologies continues to advance our understanding of complex microbial communities. It has already had a major impact on life science and is a well-recognised tool in forensic sciences and is being widely used to study various diseases including cancer, TB, AIDS etc It has already had a major impact on life science research by speeding up the sequencing of genomes and is a monumental technique of molecular biology with the gene amplification market predicted to grow to $2.2 billion by 2017. However, despite their great specificity, early PCR techniques were not suitable for routine use by the clinical diagnostic community due to the inability to effectively implement them on a large scale.
In a new development Dr Vikash Bhardwaj, a scientist from Lovely Professional University has researched on DNA more and developed a new PD-PCR(‘Parallel DNA Polymerase Chain Reaction’) technique which claims synthesis of a new DNA having polarity opposite to the template used. A step further than Nobel Prize winning scientific techniques of the 20th century, it was published in the prestigious F1000Research Journal of UK. Talking about his discovery Bhardwaj shares: “It took nearly 85 years to universally accept DNA as a key molecule which contains all information necessary for the sustenance of an organism. Initially, it was really challenging for me to think beyond conventional knowledge. This research will be helpful in tracing all errors hitherto committed and their analysis will further help researchers to find better solutions to eradicate fatal diseases like cancer, tuberculosis, AIDS etc.
Native of Kanjhwala, a rural village in Delhi, Bhardwaj has been a bright student right since childhood when he cleared Jawahar Navodaya Vidyalya (JNV) entrance exam and got free education at JNV Mungeshpur Delhi upto class 12. He completed his graduation from University of Delhi and qualified for JNU-DBT combined biotechnology entrance exam. He was selected for M.Sc. Industrial Biotechnology programme of Sardar Patel University Gujarat where he topped with first class distinction. It was during his post graduation study, that he cleared UGC-CSIR NET, DBT JRF, ICMR JRF and GATE Exam (97.2 percentile). In 2006, he applied for SCMM JNU PhD exam and finally topped that completing his PhD under the guidance of Dr Chinmay Mukhopadhyay. His PhD work was awarded Best Poster Award at SBC conference held at IISC Banglore in 2010. Bhardwaj joined Sardar Patel University Gujarat as Asst. Prof/ Research Scientist post his PhD and is currently teaching biotechnology (Molecular Biology) to M.Sc students at Lovely Professional University along with his routine research interest on “Parallel stranded DNA”. He has spent eight years so far in molecular biology research.
So how did he get the idea for this discovery? “I was aware about one of research published by scientists from Paris where they tried to prove parallel DNA synthesis but were finally unsuccessful to accomplish it. It was challenging to find out the reasons behind that, since they did not discuss much details. In collaboration with Kulbhushan Sharma (who is also a hardcore molecular biologist involved in exploring various cell signalling pathways at INMAS,DRDO), I accepted the challenge and started proving parallel DNA synthesis using many different possible ways,” he reveals. It was during one of the reactions that they were surprised to see the amplification of same size PCR product as similar to control reaction. “It was not a direct approach to develop this reaction. We used few combinations of primers in different ways but were surprised to see a similar reaction as a control reaction. Then we re-analysed our whole strategy, checked the pair of primers used and developed PD-PCR this way,” he adds.
In a conventional PCR reaction, oligonucleotide primers binds to a template DNA in an antiparallel complementary ways and DNA is amplified as it is. In a PD-PCR reaction, Bhardwaj along with his fellow researcher has shown that using a single stranded DNA as template, even parallel complementary primer can initiate a DNA synthesis reaction and finally a different but related product can be synthesised. Thus two PCR products can be synthesised from a single stranded DNA, one by conventional PCR and another by PD-PCR. So when the discover of PCR can have far reaching applications, the discovery of this kind of DNA amplification would definitely be a gamechanger in molecular biology.
“One main application of our work can be to resolve errors observed in other molecular techniques (Southern/ northern blotting, microarrays, RNAi technique, and even PCR), which all are based on complementary binding of two strands of nucleic acid. Various scientists have proposed numerous technical reasons behind non-specific errors of these techniques, but none of the scientific reports have taken into consideration the parallel complementary binding of two strands of DNA. We strongly believe that many molecular techniques developed so far can be further improved in the future considering both parallel and antiparallel complementarity of DNA. We strongly believe that researchers will understand the significance of our work, which will lead to a clearer understanding of the various biological phenomenons. Higher accuracy in molecular biology research worldwide will lead to better solutions for mankind’s problems,” stresses Bhardwaj.
It is believed that most of our DNA contains ‘Junk DNA’ and over the years, researchers have found evidence to suggest that junk DNA may provide some form of functional activity. “We believe strongly that nature do not create anything ‘Junk.’ Hopefully our study will help in better understanding of Junk DNA also. Initially, we are exploring various genomes and have successfully developed a manual strategy to find sequences in DNA which can form both parallel and antiparallel stranded DNA. Our initial finding says that most of genes contain DNA sequences which have the potentiality to form both parallel and antiparallel DNA. It’s again a challenge for us to find out the function of those sequences in future,” he opines.
Bhardwaj currently does not have any grants to carry out the research further, but he will be applying to various funding agencies for the same. When asked if Indian R&D is suffereing due to few grants and the role of academic institutions in the same, he agrees that while it is true that researchers in West receive huge grants to work on their ideas, however, Indian scientists are also getting huge grants. However, these aren’t available to young researchers. “I feel funding agencies can make a difference by understanding that many great discoveries in science were by young people. They must be liberal to adopt and fund young researchers,” he says.
Congratulating Dr Bhardwaj, LPU Chancellor Ashok Mittal offers another perspective, “Government funding agencies should be liberal in adopting researchers to stop brain drain. Research work is a never ending process which brings more and more of positive results, as such funding agencies should not limit a researcher right from the beginning about the results. Sometimes, research results can be diversified also. However, I feel that researchers should lay determined stress on the not traversed or unsuccessful areas to provide possible novel results.”
Meanwhile, Bhardwaj has been invited to give a talk in a PCR Congress which is going to held in San Diego USA where approximately 175 biotechnology scientists from top of the world institutions will be taking part. He is confident of making an inpact on the scientific and research community and hopes to collaborate with them in the future.
“This research work is my leap towards the aim of my working place LPU to get included in the list of top 200 universities of the world by the year 2025. I am now really excited and looking for grants to carry out further research,” he concludes.
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