Apr 20th, 1:00 PM - 4:00 PM
Enhancing PCR Amplification From Diluted DNA Samples Using Experimental Formulas from Biomatrica
Very dilute DNA samples present problem for forensic analysts. These samples often degrade, and their low concentration makes DNA amplification difficult or impossible. Recently a new compound was produced (PCRboost, Biomatrica) based on molecules found in water bears (tardigrades, see image). These tiny remarkable animals can live up to 120 years without water, survive temperatures from close to absolute zero to 151°C (303°F), and recover after being exposed to the vacuum of outer space. Dr. Hampikian’s laboratory at Boise State University is working with Biomatrica to assess new formulations of their product aimed at enhancing forensic DNA analysis. In this study, I show the results of DNA tests done on my own diluted DNA with several experimental formulations from Biomatrica. DNA was extracted using the QIAmp DNA Mini kit (QIAGEN), from cheek swabs and then amplified by PCR (polymerase chain reaction). Using quantitative realtime PCR the amount of quantifiable human DNA was accurately determined. The quantified DNA was then diluted down to 1.0 picogram/microliter. The diluted DNA was then used to test amplification effectiveness using ABgene standard Taq polymerase with and without the experimental products (Biomatrica) added to the reaction mixture. The amount of DNA amplified with and without each of three experimental products was compared by first measuring the amount of PCR product, and then evaluating the quality of sequencing reactions for each experimental compound.