Hafsa Ahmed Khan

APPLICATION OF EMULSION PCR FOR MULTIPLE SNP ANALYSIS

Emulsion PCR is a well-known technique allowing for the creation of in-vitro compartments. This compartmentalization helps in reducing PCR biasness by making it possible for more and all kinds of DNA templates to find their way into a polymerase chain reaction. It also works to reduce intermediate product formation. The water-in-oil emulsion technique gives rise to droplets of enough stability and strength that each one of them ends up acting as a complete PCR itself. In this study, we explored the potentials of using an easy mix of Mineral Oil based emulsion for ePCR under different conditions. For further exploration in this avenue, we developed a novel formula employing Coconut Oil as an alternative to Mineral Oil in emulsions for ePCR. Our studies concluded that ePCR does infact result in performing the polymerase chain reaction more efficiently than a conventional PCR. The percentage increase trends for the total amplified DNA product obtained at the end of different ePCR runs were observed quantitatively using Image J analysis and densitometry calculations.

Future Prospects:

This technology has great implications to be used in multiplex and assembly PCR techniques as it allows for different sized DNA templates to undergo processing simultaneously, resulting in a very efficient way of reducing time and cost for laboratory protocols and POC diagnostic techniques.