Improving current methodology used in the forensic analysis of bisulfite modified DNA samples Haley Ecker, Johnisa Walcott, Deborah Silva Chemistry Department, Hofstra University, Hempstead-NY, 11549, USA
Introduction
Results
Studying DNA methylation patterns is advantageous for the forensic field as it can be used to identify the type of tissue or fluid at a crime scene, determine the sex of the sample donor, estimate the age of the sample donor, and distinguish between monozygotic twins. Bisulfite modification is a method often used in DNA methylation. This is a harsh process that changes the chemical structure of the DNA and can damage the strands, making it not ideal for forensic samples. The main goal of this research was to modify the current methodology used in bisulfite modification and subsequent PCR to repair damaged DNA and to improve data generation and the downstream analysis of results.
The protocol using the EZ DNA methylation-lightning kit coupled with the added repair enzymes to the PCR process presented the best performance and higher yield of PCR products.
Methodology Modified PCR Epitect kit
Nanodrop One Standard PCR
Sample Modified PCR EZ kit
Nanodrop One Standard PCR
The results obtained in this initial study show that it is possible to improve the quality of DNA samples after going through a harsh chemical modification, which will then provide better results in DNA amplification and other downstream methods and analysis.