a)Downstream processing: for the purpose of RT-PCR IVD kits, Taq DNA polymerase is needed. I optimized the buffer and chemical component to improve its activity. Now I am working on MasterMix formation for RT-PCR kit. b) Cost cutting, previously we were using Maxi kit for plasmids from Qiagen to isolate bulk DNA for DNA ladder formation. I ordered the maxi prep column only and optimized buffers for the column elution of plasmids to get the clean band in DNA ladder. Previously we were buying the 6X DNA dye from outside, I made our own DNA dye, now cost goes down further.C) Upper band (undesired) was coming to 50 bp and 1 kb DNA ladder, I trouble shoot the problem, changes the DNA extraction buffer, now the band quality is upto the mark without extra upper band.

a) Downstream processing of Bst DNA polymerase, Reverse transcriptase and Taq DNA polymerase for the purpose of LAMP, RT-LAMP and RT-PCR kit formation. b) Technical support to the customers, demonstrating the kits to customers and troubleshooting their experiments.

Worked on cloning of corona genome assembly in multicopy vector inside Saccharomyces cerevisiae.

Worked on the Enzymes Expression and purifications.

Worked to find a novel easy and affordable method to develop a kit for detection of toxin inside milk.

I worked on human coat protein BBS5 (a subunit of BBSome) protein . I needed BBS5 protein in soluble, refolded and mono-disperse form. The cloning of BBS5 gene into various expression vectors for E. coli was done to get the soluble expression of BBS5 proteins. After examining various growth conditions for expression of BBS5 in E. coli, it was found that BBS5 protein is getting inside the inclusion bodies. Different refolding buffer conditions were applied to BBS5 inclusion bodies. Besides this, co-expression of other proteins like Maltose binding protein/ Glutathione S transferase proteins was also done to get soluble folded protein. Subsequently, truncation library was created and clones were screened for their expression in soluble form inside cytosol. To ease the selection process, fluorescent proteins were tagged to the C –terminus end of the protein. Hence, clones with soluble protein expression were visualized via exposure to the UV/visible ray. I found many clones of truncated size showing fluorescent colour when given exposure of UV/visible light. These clones were giving soluble truncated BBS5 protein in their cytoplasm.

I worked on an industry sponsored project entitled “Fourth generation clot bluster” at Institute of Microbial Technology , Chandigarh. Streptokinase (SK) is a thrombolytic protein that acts as a co-factor for human plasmin (ogen) (HPG/HPN). To improve its activity, initially I did some analysis using bioinformatic tools like to find the flexible/ disordered regions in SK. Different bioinformatics tools gave different predictions. Hence, after analyzing those results I chose the region which was predicted by all the tools. Subsequently, I did saturation mutagenesis of those residues and selected few residue on the basis of gain in activities.