We at Indigenèse Biotechnologies have molecular biology training and education as one of our principal goals. As a part of this mandate we organize short workshops with a focus on giving hands-on training tothe participants. The first such workshop was organized in December 2006 for college and high school teachers in basic molecular biology techniques. The second one in the series was organized for a group predominated by physicians specializing in clinical microbiology. This workshop was conducted at, and in collaboration with, the Nizam's Institute of Medical Sciences, in Hyderabad on 13-14 July, 2007. 

 

The workshop began on 13 July 2007 with an inaugural function. Dr.G.Lakshmi, Head and Professor of Microbiology who spearheaded and coordinated the workshop introduced the workshop. Dr. Dharmarakshak who was deputing for the Director of the Institute presided over the inaugural session. The common theme of their introductory remarks was that DNA based molecular tools are becoming an integral part of modern clinical settings and that there is a great need for training clinical personnel in these techniques. Dr Anil Kumar Challa gave an hour long presentation on the brief history of DNA Science and its impact on medical science.  Beginning with the history of classical genetics, he talked about the molecular biology revolution and the rise of recombinant DNA (rDNA) technology.  He then gave a brief overview of the impact of rDNA Technology on clinical medicine, especially on microbiology. 

 

The hands-on sessions started off with activities on DNA extraction from human cheek cells followed by isolation of plasmid DNA from recombinant bacteria. Ms. G. Ramya conducted this session and started off with the conceptual basis for isolation of DNA from cells. Participants used their cheek cells to extract human genomic DNA. A simple protocol of lysing the cells open with detergent solution, and precipitating the DNA with cold isopropanol was used.  All the participants were able to visualize their DNA. They were excited to see the white strands of DNA slowly precipitating, upon the addition of alcohol. This was followed by the isolation of plasmid DNA from bacterial cells using an alkaline lysis protocol. 

 

The post-lunch session began with an introduction to restriction enzymes, a brief history, their function and role in modern molecular biological laboratories by Ms. Neha Joshi. The participants were taken through the history of phage restriction-modification system discovered in the 1950s and how those initial genetic experiments led to the discovery of the site specific DNA endonucleases as invaluable tools for molecular biology. The discussion was followed by an activity wherein the participants did a double digest of the plasmid (using two restriction endonucleases) that they had isolated earlier.

 

Day two started off with an introduction to Polymerase Chain Reaction by Ms. Ambika Srinivasan. In addition to discussing the underlying concept of PCR, a detailed description of the components, factors and conditions making the PCR possible was given. Each participant set up their own PCR using a recombinant plasmid DNA as a template to amplify an 800 bp fragment inserted in the plasmid. Introduction to Agarose Gel Electrophoresis and its utility in separating DNA fragments was given by Ms. G.Ramya. The participants then loaded the products of restriction digestion reaction from day one.  Some enthusiastic participants also loaded food colors for practice. A discussion on the applications of PCR ensued while the agarose gel was running. Due to the wide usage of PCR technologies in clinical diagnosis many of the participants actively contributed to the discussion. The greater interest in the discussion was due to the  immense popularity of the technique in many fields. Some of the participants expressed interest in setting up PCR labs at their respective institutions. Dr. G Lakshmi contributed to the discussion by sharing her experiences and insights in setting up laboratories capable of performing these molecular techniques.  On the point of use of PCR in GMO testing, Dr. Shyamala (Blood Bank, Apollo Hospitals) raised questions about why there is fear of GMOs in people. The participants had questions on DNA fingerprinting and 16s rRNA typing (Dr MVS Subbalakshmi).

Dr. M.U.R.Naidu, Dean of NIMS, who visited the laboratory during the workshop, was glad that this hands-on workshop was happening and that he would be interested in similar workshops for senior physicians on Sundays to enable them to participate as well.

The post-lunch session began with an introduction to bioinformatics. Dr Anil Kumar Challa briefly described the databases at NCBI and engaged the participants in doing an exercise in literature search using PubMed followed by another exercise on retrieving a nucleotide sequence from the Entrez database and doing a BLAST analysis.

During this discussion the IB team loaded the PCR amplification products on to agarose gels to perform electrophoretic separation.  Subsequently the participants viewed the gels and observed the DNA bands in their restriction digestion reactions and the PCR amplified products. While the restriction enzyme digestion reaction products were not very well observed on the gels, robust bands of DNA were observed in the PCR amplifications. The participants were excited to visualize their PCR amplified products on an agarose gel under UV illumination.

 

After the laboratory sessions ended, Dr. G Lakshmi invited the participants to share their reflections about the workshop. There was a consensus amongst the participants that this was a unique workshop and they learnt a lot, not just in terms of the theory but also doing every technique with their own hands.  Most of the participants agreed that the workshop fee was well worth the experience.  Some of the participants opined that conducting more of these hands-on workshops would enable postgraduate microbiology students to get an insight into modern molecular biology techniques.  And, this would potentially result in effective combination of clinical microbiology with molecular tools towards improving the quality of research and diagnosis.  They also wanted to know more about the clinical applications of these techniques.  A couple of participants asked what would be the next level of the workshop and that we should plan for the next one. Towards the end, Dr.Challa was asked by Dr. G Lakshmi to give away the workshop certificates to the participants.

This workshop became a reality with the help of many people at NIMS. Dr. Nayana Joshi was the pivot who introduced Indigenèse to Dr. G Lakshmi and championed the training mandate of Indigenèse. Dr. Lakshmi and Dr. Nayana Joshi were enthusiastic from the beginning and gave a lot of enthusiastic support. Dr. Lakshmi’s team, especially Dr. T Sudha, stood behind the scenes and took care of the logistics of the workshop. The IB team thanks all the members of the NIMS team for the success of the workshop.