Internal Validation of Reduced Polymerase Chain Reaction Volume for the Amplification of Nonquantified DNA Extracts from Reference Samples

Iqbal, Muhammad Usman; Butt, Muhammad Ahsan; Siddique, Nasir; Ahmad, Qazi Laeeque; Ahmad, Sajjad; Amjad, Muhammad; Tahir, Mohammad Ashraf

doi:10.1089/forensic.2021.0007

Forensic Genomics

2021

DOI: 10.1089/forensic.2021.0007

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Internal Validation of Reduced Polymerase Chain Reaction Volume for the Amplification of Nonquantified DNA Extracts from Reference Samples

Muhammad Usman Iqbal¹,

Muhammad Ahsan Butt²,

Nasir Siddique³

et al.

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2024

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DNA typing of cyanoacrylate fumed latent fingerprints using GlobalFiler™ and ForenSeq™ Signature Prep kits

Bodnar,

Smith,

Alsharji

et al. 2024

Journal of Forensic Sciences

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DNA typing of latent fingerprints is highly desirable to increase chances of individualization. We recovered DNA from Cyanoacrylate (CA) fumed fingerprints and used both GlobalFiler™ and ForenSeq™ DNA Signature Prep kits for DNA typing. For GlobalFiler™, samples were processed using a protocol modified for Low Template (LT)‐DNA samples (half‐volume reactions, 30 cycles) while for ForenSeq™ DNA Signature Prep, samples were processed using a standard protocol and fluorometer‐based library quantitation. We evaluated genotyping success and quality of profiles in terms of completeness, Peak Height Ratio/Allele Coverage Ratio, presence of PCR artifacts and drop‐in alleles. With GlobalFiler™, average autosomal STR (aSTR) profile completeness was 44.4% with 2–20 pg, 54.3% with 22–60 pg, and 95% with 64–250 pg DNA input. CODIS uploadable profiles were obtained in 2/10, 3/11, and 11/12 samples in these ranges. With ForenSeq™ DNA Signature Prep, average aSTR profile completeness was 19.7% with 1–20 pg and 45.2% with 22–47 pg but increased to 78.3% with 68–122 pg and 86.7% with 618–1000 pg DNA input. Uploadable profiles were obtained in 0/12, 4/11, 4/7, and 3/3 samples for these ranges. Results show very high sensitivity using both kits. Half‐volume reactions and 30 cycles had minimal negative effect on Globalfiler™ profile quality, providing support for wider use after validation experiments to routinely improve results from LT samples. A standard protocol for the ForenSeq™ DNA Signature Prep kit was also highly successful with LT DNA obtained from CA‐fumed fingerprints with additional information from isometric STR alleles and other markers.

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DNA typing of cyanoacrylate fumed latent fingerprints using GlobalFiler™ and ForenSeq™ Signature Prep kits

Bodnar,

Smith,

Alsharji

et al. 2024

Journal of Forensic Sciences

View full text Add to dashboard Cite

show abstract

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Internal Validation of Reduced Polymerase Chain Reaction Volume for the Amplification of Nonquantified DNA Extracts from Reference Samples

Cited by 1 publication

References 3 publications

DNA typing of cyanoacrylate fumed latent fingerprints using GlobalFiler™ and ForenSeq™ Signature Prep kits

DNA typing of cyanoacrylate fumed latent fingerprints using GlobalFiler™ and ForenSeq™ Signature Prep kits

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