Effects of sampling, DNA amount and Reduced volume of AmpFLSTR Identifiler Plus kit on STR profile

Abstract

This research showed two examined blood sampling option, 20 finger-prick and 20 four-month freezers venous blood and its effect on the recovery of isolated DNA preserved on FTA Cards. also, to reduce the cost of DNA profile, we estimated kit efficiency using a reduced volume of PCR reaction (12.5μL) with different DNA concentration. A prepare series of DNA dilution (0.01, 0.1, 0.5, 1 and 10 ng/ μL) to amplified DNA with two volume 12.5 and 25 μL of AmpFSTR® Profiler Plus™ Kit. Our research observed that sampling methods and storage time led to decreased DNA. The full and half PCR volume rection was produced a complete DNA profile with both 0.5 and 1 μL DNA concentration. The loss of DNA profile was detected in samples with 0.01 μL of DNA concentrations, while 0.1 μL of DNA concentration was produced partial DNA profile. The optimal DNA quantity which yielded full DNA profile were 0.5 and 1 ng/ μL with both 12.5 and 25 μL volume of PCR. Generally, our data indicated that the peak height of STR loci in histogram results, lowering progressively with decreasing of DNA amounts. Accordingly, this modified method decreases the cost of DNA profile to half, while maintaining its performance.