A single direct amplification method for forensic casework references on a variety of substrates

Chong, Mavis Date; Wallin, Jeanette M.

doi:10.1016/j.fsir.2022.100260

Cited by 1 publication

(1 citation statement)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The findings are consistent with that of previous studies [21, 22] that reported about reduction in PCR volumes, whereby purified samples often revealed more complete profiles when compared with that of direct amplification. One possible explanation for this observation is due to the increased concentration of inhibitors following the reduction of PCR reaction volume [21, 23]. Considering that all the purified samples amplified using the 10.0 μL PCR reaction volume (60% reduction) consistently revealed complete DNA profiles, such sample type and PCR reaction volume were maintained for the subsequent optimization steps.…”

Section: Resultsmentioning

confidence: 99%

Internal validation of reduced PCR reaction volume of the Qiagen Investigator® Argus X‐12 QS Kit from blood samples on FTA cards

Alwi,

Mahat,

Mohd Salleh

et al. 2023

Journal of Forensic Sciences

View full text Add to dashboard Cite

The onus of proof in criminal cases is beyond any reasonable doubt, and the issue on the lack of complete internal validation data can be manipulated when it comes to justifying the validity and reliability of the X‐chromosomal short tandem repeats analysis for court representation. Therefore, this research evaluated the efficiency of the optimized 60% reduced volumes for polymerase chain reaction (PCR) amplification using the Qiagen Investigator® Argus X‐12 QS Kit, as well as the capillary electrophoresis (CE) sample preparation for blood samples on Flinder's Technology Associates (FTA) cards. Good‐quality DNA profile (3000–12,000 RFU) from the purified blood sample on FTA card (1.2 mm) were obtained using the optimized PCR (10.0 μL of PCR reaction volume and 21 cycles) and CE (9.0 μL Hi‐Di™ Formamide and 0.3 μL DNA Size Standard 550 [BTO] and 27 s injection time) conditions. The analytical and stochastic thresholds were 100 and 200 RFU, respectively. Hence, the internal validation data supported the use of the optimized 60% reduced PCR amplification reaction volume of the Qiagen Investigator® Argus X‐12 QS Kit as well as the CE sample preparation for producing reliable DNA profiles that comply with the quality assurance standards for forensic DNA testing laboratories, while optimizing the analytical cost.

show abstract

Section: Resultsmentioning

confidence: 99%