DNA sequencing using hybridization: In the project we are aiming to sequence small strands of DNA using single molecule fluorescence technique. Sequencing technique has come a long way from early steps to next generation. However, for DNA strands containing lesser number of nucleotides for example 5-15 poses challenges. We take the approach of DNA hybridization, where complementarity in sequences controls the rate of hybridization. From the comparative rate information between DNA strands containing complete and partial complementarity we aim to identify individual bases in DNA strands. Using total internal reflection fluorescence microscopy we measure fluorescence signal due to hybridization where individual strands contain FRET pair. Pairing it with microfluidics and machine learning we aim to achieve high throughout sequencing of DNA.
Deciphering the mechanism of force-dissemination during hearing: Mammalian hearing is a very versatile system orchestrated by plethora of molecules interconnected in intricate pathways. However, the inquisitive action is controlled by two non-classical Cadherin family proteins Cadherin 23 and Protocadherin 15. They form the tip-link connecting vertically neighbouring stereocilia in hair cell in the inner ear. Sound induced mechanical vibration creates oscillation in the stereocilia which opens up the ion channels present at the bottom of tip-link. Upon opening ions from extracellular medium flows through the ion channels and change the cell polarity which is being sensed by the afferent auditory nerve. We are trying to understand the force-sensation mechanism of tip-link and how it can withstand huge ranges of forces of magnitude 2-80 pN coming from sound of 20 Hz-20 kHz and 0-120 dB. For this project we utilized AFM based force measurements and TIRF based single molecule fluorescence techniques.