Validation of reduced volume VeriFiler™ Express PCR Amplification Kit for buccal swab samples extracted using Prep-n-Go™ Buffer.

ABSTRACT: The efficiency of reduced volume PCR amplification was studied using the VeriFiler™ Express PCR Amplification Kit. Full (25 μL) and reduced (5 μL) volumes were tested in parallel to identify any differences in template DNA sensitivity and other electropherogram parameters. Both volumes produced full DNA profiles down to 0.08 ng/μL DNA concentration at 26 PCR cycles; however, reduced volume produced higher peak heights due to increased signal intensities. Significant difference (p-value ≤ 0.05) in heterozygote peak height ratios was observed between both volumes, where the reduced volume threshold was lowered to 0.6 to accommodate all data points. However, no significant difference (p-value > 0.05) was identified in the stutter ratios between both volumes. The analytical threshold for reduc

SUBMITTER: Perry J 

PROVIDER: S-EPMC9546378 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature