Short amplicon STR multiplex for stain typing

Wiegand, Peter; Klein, Rachel Herndon; Braunschweiger, G.; Hohoff, Carsten; Brinkmann, B.

doi:10.1007/s00414-005-0048-8

Cited by 35 publications

(24 citation statements)

References 14 publications

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“…The extracts were then subsequently purified with silica suspension (Dianova, Hamburg, Germany) as described in ref. [2]. Besides, 10 mL of ProteinaseK (QIAGEN, Hilden, Germany) and 20 mL of 1 M DTT (Sigma-Aldrich) were added to the lysis buffer in the morning and evening.…”

Section: Methodsmentioning

confidence: 99%

“…PCRs were performed using the multiplex according to [2] plus the system SE33 with shortened amplicons. The primer sequences for the locus SE33 were the following: P1-JOE:…”

Section: Methodsmentioning

confidence: 99%

“…Over the last few years some laboratories have developed improved extraction methods and miniSTR kits (short-amplicon PCR) to increase the typing chance of highly degraded and weak stains [1][2][3][4][5][6]. We present an extraction method which can be used successfully for biological stains, like telogen hair, with little and highly degraded DNA.…”

Section: Introductionmentioning

confidence: 98%

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Improved STR typing of telogen hair root and hair shaft DNA

et al. 2007

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Today the STR typing of telogen hair and hair shafts is regarded as a challenge. The small DNA quantity in the hair is highly degraded. Another problem are PCR inhibitors in the hair. In particular hair pigments, the melanins, are known to inhibit PCR. Hairs are exposed to sunlight and partly to chemical oxidation processes, which make them even more difficult to analyze. To increase the chances of a correct typing of hair, the small amount of DNA must be successfully isolated and the inhibitors have to be removed or neutralized. Furthermore, miniSTR typing improves the analysis of stains with degraded DNA like it is the case with hair. We introduce a nonorganic extraction method and in addition a miniSTR concept which is promising in typing stains with little and degraded DNA, especially hairs. The miniSTR concept including five database STRs (SE33, VWA, TH01, FGA, D3S1358) and the gender typing system Amelogenin was optimized for the amplification of hair DNA. Compared to commercial STR kits, this approach resulted in considerably higher success rates.

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Section: Methodsmentioning

confidence: 99%

“…PCRs were performed using the multiplex according to [2] plus the system SE33 with shortened amplicons. The primer sequences for the locus SE33 were the following: P1-JOE:…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 98%

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Improved STR typing of telogen hair root and hair shaft DNA

et al. 2007

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“…However, both a very close correlation between the quantity and quality of the polymorphic loci present in an STR system, and the degree of accuracy of the methodology, are evident. Thus, the greater the number of STRs analyzed, the greater would be the resolving power of the study (Wiegand et al, 2006). This study aimed to develop a new STR multiplex system comprising CODIS and non-CODIS highly polymorphic loci for simultaneous analysis, and to assess its efficiency for forensic application.…”

Section: Introductionmentioning

confidence: 99%

“…According to Wiegand et al (2006), several real situations of high analytical complexity require a molecular arsenal focused on the power of individual discrimination. In these situations, the application of methods that increase the power of discrimination corroborates the issuance of conclusive opinions.…”

Section: Introductionmentioning

confidence: 99%

Development of a polymorphic short tandem repeat locus multiplex system for efficient human identification

Rodovalho

Rodrigues

Santos³

et al. 2017

Genet. Mol. Res.

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ABSTRACT. This study aimed to develop a short tandem repeat (STR) multiplex system, made up of 22 highly informative STR loci, for application in forensic genetics. The system comprised 21 polymorphic autosomal loci (D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, 2 R.G. Rodovalho et al.Genetics and Molecular Research 16 (2): gmr16029557 D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX, FGA, D2S441, D17S1301, D19S433, D18S853, D20S482, and D14S1434) and the amelogenin gene locus. Strategies were developed to overcome the challenges involved in creating a multiplex system. Based on the literature and available databases, the STR loci were selected with the aim to obtain discriminatory markers, and followed specific criteria for this purpose. Primers were projected using the Primer3 software, and AutoDimer was used to evaluate possible interactions between them. The 22 selected STR loci were validated individually and jointly, both to assess their sensitivity and to test the efficiency of the multiplex system. Statistical analyses were based on the genetic data of 450 unrelated individuals living in the State of Goiás, thus allowing the establishment of the parameters necessary to use this system. A total of 239 alleles were detected for the 22 loci in the set, allowing for a probability of identity of 4.23 x 10 -25 to be obtained. The combined power of discrimination was 0.999999999999999999999999 and the combined power of exclusion was 0.99999. Upon complete validation of the entire system, this multiplex assay was considered to be a powerful tool for application in human identification by DNA analysis.

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Comparison of massively parallel sequencing to capillary electrophoresis for short tandem repeat genotyping of trace DNA

Dierig,

Kunz,

Wiegand

2023

Electrophoresis

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In forensic genetics, massively parallel sequencing (MPS) offers several advantages over the current golden standard, capillary electrophoresis (CE): additional sequence information, shorter amplicon lengths, and the simultaneous analysis of many markers. These benefits result in a reduced number of reactions necessary while improving the amount of data obtained, thereby conserving valuable sample extracts. This proves particularly advantageous for the analysis of trace DNA. This study assessed the suitability of MPS for short tandem repeat (STR) typing of low template samples compared with results obtained through CE. The MPS genotypes showed higher concordance to reference genotypes, with donor alleles being more frequently assigned to be the major contributor, meeting the requirements for database entry. However, the MPS workflow is more time‐consuming and associated with higher costs.

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Short amplicon STR multiplex for stain typing

Cited by 35 publications

References 14 publications

Improved STR typing of telogen hair root and hair shaft DNA

Improved STR typing of telogen hair root and hair shaft DNA

Development of a polymorphic short tandem repeat locus multiplex system for efficient human identification

Comparison of massively parallel sequencing to capillary electrophoresis for short tandem repeat genotyping of trace DNA

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