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Developmental validation of the STRscan-17LC kit: a 6 Dye STR kit enhanced stability and ability to detect degraded samples

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Abstract

Genotyping of short tandem repeat (STR) markers is the basic method of forensic science. Enhanced technologies are needed to meet the requirements of databasing and casework samples. The STRscan-17LC kit is a 6 Dye STR kit which amplifies 16 STR loci: D3S1358, TPOX, D16S539, vWA, D2S1338, CSF1PO, D19S433, D7S820, FGA, D8S1179, D5S818, D13S317, D18S51, TH01, D12S391, and D21S11 and the sex-determinant locus amelogenin. This kit is designed for better tolerance to PCR inhibitors and analysis of mildly degraded samples with all fragments smaller than 330 bases. In this study, the STRscan-17LC kit is validated according to the SWGDAM (Scientific Working Group on DNA Analysis Methods) guidelines, including PCR-based studies, sensitivity, precision and accuracy, inhibitors, species specificity, DNA mixture studies, population, and concordance studies. The validation results suggest that the STRscan-17LC kit is a useful tool for forensic application.

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Funding

This project was supported by the National Natural Science Foundation of China (NSFC, No. 81871533), the Natural Science Foundation of Hunan Province (No. 2020JJ4779), and China Postdoctoral Science Foundation (No. 2020M682591).

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Author notes

  1. Bowei Jiang and Wei He contributed equally to this work and should be considered as co-first authors.

Authors and Affiliations

  1. The First Research Institute of the Ministry of Public Security P.R.C, Beijing, People’s Republic of China

    Chuan Jin

  2. Department of Forensic Science, School of Basic Medical Sciences, Central South University, No172. Tongzipo Road, Changsha, 410013, Hunan Province, People’s Republic of China

    Ying Liu, Dan Wen, Chudong Wang, Moutanou Modeste Judes ZEYE, Jienan Li & Lagabaiyila Zha

Authors
  1. Bowei Jiang
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  2. Wei He
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Correspondence to Lagabaiyila Zha.

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Supplementary information

Fig. S1

Electropherograms of control DNA 2800 M amplified with different PCR reaction volumes (5 μL, 10 μL and 20 μL). (JPG 634 kb)

Fig. S2

Effect of 1x, 0.75x, 0.5x buffer consentrations and 1x, 0.75x, 0.5x Primer Mix consentrations on the STRscan-17LC kit. (JPG 490 kb)

Fig. S3

Effect of various cycle numbers (24, 26, 28, 30 and 32) on the STRscan-17LC kit. (JPG 370 kb)

Fig. S4

Effect of various annealing temperature (55, 57, 59, 61 and 63 °C) on the STRscan-17LC kit. (JPG 406 kb)

Fig. S5

Effects of shortening the final extension time after normal thermal cycling for 1 ng 2800 M. The red box shows a minus-A shoulder peaks at the CSF1PO locus. (JPG 242 kb)

Fig. S6

Representative electropherograms from species specificity studies. (JPG 1008 kb)

Fig. S7

Electropherograms from sensitivity study samples were tested with a serial dilution of following template amounts: 1 ng, 500 pg, 250 pg, 125 pg, 62.5 pg, and 31.25 pg. (JPG 1191 kb)

Fig. S8

Precision across 32 injections of the STRscan-17LC kit allelic ladder mix performed on 3130xl Genetic Analyzer. (JPG 333 kb)

Fig. S9

Male/female mixtures (9948 and 9947A human genomic DNA) were prepared and tested with the ratio of 1:0, 0:1, 1:1, 1:4, 1:9, and 1:19. (JPG 428 kb)

Fig. S10

Hematin were tested with the STRscan-17LC kit. When added nothing, full profiles were obtained. The experiments were carried out with hematin of 0uM, 50uM, 100uM, 150uMand 200uM, respectively. With the increase of hematin concentration, the dropout of alleles increased.When the concentration of hematin was raised to 200uM, complete dropout occurred. (JPG 364 kb)

Fig. S11

Humic acid were tested with the STRscan-17LC kit. When added nothing, full profiles were obtained. The experiments were carried out with humic acid of 0 ng/ul, 100 ng/ul, 200 ng/ul, 300 ng/ul, 400 ng/ul and 500 ng/ul, respectively. With the increase of humic acid concentration, the dropout of alleles increased. When the concentration of humic acid was raised to 500 ng/ul, complete dropout occurred. (JPG 437 kb)

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Jiang, B., He, W., Jin, C. et al. Developmental validation of the STRscan-17LC kit: a 6 Dye STR kit enhanced stability and ability to detect degraded samples. Int J Legal Med 135, 431–440 (2021). https://doi.org/10.1007/s00414-020-02490-y

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  • DOI: https://doi.org/10.1007/s00414-020-02490-y

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