Reduced PCR cycling time amplification using AmpFℓSTR ®  Identifiler ®  kit

Iyavoo, Sasitaran; Wolejko, Agata; Furmanczyk, Dagmara; Graham, Richard; Myers, R. J.; Haizel, Thomas

doi:10.1016/j.fsigss.2015.09.113

Cited by 6 publications

(3 citation statements)

References 5 publications

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“…Increasing the speed of the PCR amplification step can be done in a variety of ways. Some groups modify commercially available kits and their protocols, while others create entirely new multiplexes specially designed for fast amplification. , Gibson-Daw et al used a 7-locus multiplex on a high speed thermocycler and a rapid polymerase to achieve a multiplex amplification in 6.5 min . Lower total volumes have been shown to help reduce the time of heating and cooling of the sample, reducing amplification time by 56–73% .…”

Section: Genotyping Methods Using Short Tandem Repeatsmentioning

confidence: 99%

Forensic DNA Analysis

McCord

Gauthier

Cho³

et al. 2018

Anal. Chem.

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Section: Genotyping Methods Using Short Tandem Repeatsmentioning

confidence: 99%

Forensic DNA Analysis

McCord

Gauthier

Cho³

et al. 2018

Anal. Chem.

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“…It is becoming more common for users of these kits to validate using a lower PCR reaction mix volume to get more PCR reactions out of one kit. The validation of reduced volume PCR has been achieved with many common multiplex STR kits, for example, the PowerPlex® 16 System [1], AmpFLSTR™ Identifiler™ PCR Amplification Kit [2], and GlobalFiler™ PCR Amplification Kit [3]. Overall, reducing the PCR volume reduces cost [4] and has been found to increase the sensitivity and reduce the amount of template DNA required [1].…”

Section: Introductionmentioning

confidence: 99%

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

Perry

Munshi

Haizel³

et al. 2022

Journal of Forensic Sciences

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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 reduced volume was also determined to be 150 RFU with the presence of template DNA in PCR amplification. When the optimized reduced volume parameters were tested on DNA extracted from buccal swab samples using Prep-n-Go™ Buffer, good quality DNA profiles were produced. Overall, the reduced volume not only showed better results compared to the full volume, but also enable more samples to be processed with a PCR amplification kit, thus reduced the cost.

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“…In total 40 buccal swabs from three routine casework batches were extracted and amplified with the current accredited procedures [3,4] employing QIAamp ® DNA Mini Kit (Qiagen ® , UK) and AmpFℓSTR ® Identifiler ® PCR Amplification Kit (Thermo Fisher Scientific ® , UK) at the old premises together with the blank, positive and negative controls for each batch.…”

Section: Sample Selection Dna Extraction and Pcr Amplificationmentioning

confidence: 99%

Validation of ABI 3500xL Genetic Analyzer after decommissioned and recommissioned at new premises

Iyavoo

Draus-Barini

Haizel

2017

Forensic Science International: Genetics Supplement Series

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This validation was aimed to demonstrate that there is no effect on the Applied Biosystems ® 3500xL Genetic Analyzer (Thermo Fisher Scientific ® , UK) after decommissioned and recommissioned at new premises. Same sample sets were tested on the ABI 3500xL Genetic Analyzer at both old and new premises using current accredited procedures. The results showed genotyping concordance which means using the ABI 3500xL Genetic Analyzer at the new premises provides no risk regarding the accuracy, precision and reproducibility of the results and also showed a very good performance on sensitivity, specificity, repeatability and good profile quality based on the assessment of locus peak balance and peak height.

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Reduced PCR cycling time amplification using AmpFℓSTR ® Identifiler ® kit

Cited by 6 publications

References 5 publications

Forensic DNA Analysis

Forensic DNA Analysis

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

Validation of ABI 3500xL Genetic Analyzer after decommissioned and recommissioned at new premises

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