Improved resolution of mixed STR profiles using a fully automated differential cell lysis/DNA extraction method
Sexual assault evidence often contains sperm cells, which are typically separated from nonsperm cells using manual differential lysis procedures. The goal of this study was to evaluate the automated QIAGEN QIAcube for this purpose and to compare it to manual QIAGEN and manual organic differential me...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Oxford University Press
2020-04-01
|
| Series: | Forensic Sciences Research |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/20961790.2019.1646479 |
| _version_ | 1797720917301788672 |
|---|---|
| author | Matthew C. Goldstein Jordan O. Cox Lori B. Seman Tracey Dawson Cruz |
| author_facet | Matthew C. Goldstein Jordan O. Cox Lori B. Seman Tracey Dawson Cruz |
| author_sort | Matthew C. Goldstein |
| collection | DOAJ |
| description | Sexual assault evidence often contains sperm cells, which are typically separated from nonsperm cells using manual differential lysis procedures. The goal of this study was to evaluate the automated QIAGEN QIAcube for this purpose and to compare it to manual QIAGEN and manual organic differential methods using DNA yields and STR profile data for assessment. DNA yields were determined by qPCR, followed by multiplex STR amplification, CE analysis, and mixture interpretation. The automated method was capable of effective cell separation, producing DNA yields sufficient for STR amplification. Further, sperm fraction human:male DNA ratios from the QIAcube samples were consistently closer to the desired 1:1 and STR profiles were less likely to result in mixtures, with 6–8× fewer female alleles detected (median 1.5 alleles). Ultimately, using the QIAcube for automated differential processing of semen-containing mixtures reduces the need for downstream mixture interpretation and improves STR profile quality with substantially less hands-on time. |
| first_indexed | 2024-03-12T09:26:29Z |
| format | Article |
| id | doaj.art-86968418aef2480281cb7fc69dc4e114 |
| institution | Directory Open Access Journal |
| issn | 2096-1790 2471-1411 |
| language | English |
| last_indexed | 2024-03-12T09:26:29Z |
| publishDate | 2020-04-01 |
| publisher | Oxford University Press |
| record_format | Article |
| series | Forensic Sciences Research |
| spelling | doaj.art-86968418aef2480281cb7fc69dc4e1142023-09-02T14:10:50ZengOxford University PressForensic Sciences Research2096-17902471-14112020-04-015210611210.1080/20961790.2019.16464791646479Improved resolution of mixed STR profiles using a fully automated differential cell lysis/DNA extraction methodMatthew C. Goldstein0Jordan O. Cox1Lori B. Seman2Tracey Dawson Cruz3Virginia Commonwealth UniversityVirginia Commonwealth UniversityVirginia Commonwealth UniversityVirginia Commonwealth UniversitySexual assault evidence often contains sperm cells, which are typically separated from nonsperm cells using manual differential lysis procedures. The goal of this study was to evaluate the automated QIAGEN QIAcube for this purpose and to compare it to manual QIAGEN and manual organic differential methods using DNA yields and STR profile data for assessment. DNA yields were determined by qPCR, followed by multiplex STR amplification, CE analysis, and mixture interpretation. The automated method was capable of effective cell separation, producing DNA yields sufficient for STR amplification. Further, sperm fraction human:male DNA ratios from the QIAcube samples were consistently closer to the desired 1:1 and STR profiles were less likely to result in mixtures, with 6–8× fewer female alleles detected (median 1.5 alleles). Ultimately, using the QIAcube for automated differential processing of semen-containing mixtures reduces the need for downstream mixture interpretation and improves STR profile quality with substantially less hands-on time.http://dx.doi.org/10.1080/20961790.2019.1646479forensic sciencesforensic geneticssexual assault evidenceqiacubedifferential lysissperm lysisautomated dna extractionmixture analysis |
| spellingShingle | Matthew C. Goldstein Jordan O. Cox Lori B. Seman Tracey Dawson Cruz Improved resolution of mixed STR profiles using a fully automated differential cell lysis/DNA extraction method Forensic Sciences Research forensic sciences forensic genetics sexual assault evidence qiacube differential lysis sperm lysis automated dna extraction mixture analysis |
| title | Improved resolution of mixed STR profiles using a fully automated differential cell lysis/DNA extraction method |
| title_full | Improved resolution of mixed STR profiles using a fully automated differential cell lysis/DNA extraction method |
| title_fullStr | Improved resolution of mixed STR profiles using a fully automated differential cell lysis/DNA extraction method |
| title_full_unstemmed | Improved resolution of mixed STR profiles using a fully automated differential cell lysis/DNA extraction method |
| title_short | Improved resolution of mixed STR profiles using a fully automated differential cell lysis/DNA extraction method |
| title_sort | improved resolution of mixed str profiles using a fully automated differential cell lysis dna extraction method |
| topic | forensic sciences forensic genetics sexual assault evidence qiacube differential lysis sperm lysis automated dna extraction mixture analysis |
| url | http://dx.doi.org/10.1080/20961790.2019.1646479 |
| work_keys_str_mv | AT matthewcgoldstein improvedresolutionofmixedstrprofilesusingafullyautomateddifferentialcelllysisdnaextractionmethod AT jordanocox improvedresolutionofmixedstrprofilesusingafullyautomateddifferentialcelllysisdnaextractionmethod AT loribseman improvedresolutionofmixedstrprofilesusingafullyautomateddifferentialcelllysisdnaextractionmethod AT traceydawsoncruz improvedresolutionofmixedstrprofilesusingafullyautomateddifferentialcelllysisdnaextractionmethod |