A single direct amplification method for forensic casework references on a variety of substrates
A single direct amplification method for buccal and blood reference samples, on different substrates, containing a wide range of cellular material, was developed. This method uses the GlobalFiler™ STR kit, which was designed for amplification of previously purified and quantified DNA. Buccal samples...
| Main Authors: | , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-07-01
|
| Series: | Forensic Science International: Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2665910722000068 |
| _version_ | 1811336107505221632 |
|---|---|
| author | Mavis Date Chong Jeanette Wallin |
| author_facet | Mavis Date Chong Jeanette Wallin |
| author_sort | Mavis Date Chong |
| collection | DOAJ |
| description | A single direct amplification method for buccal and blood reference samples, on different substrates, containing a wide range of cellular material, was developed. This method uses the GlobalFiler™ STR kit, which was designed for amplification of previously purified and quantified DNA. Buccal samples on cotton swabs, Bode collectors, and Whatman® FTA® cards, and liquid blood, blood on Whatman® 903 and FTA® cards, and autopsy bloods were examined. A first pass success rate, that is, complete concordant profiles with no GeneMapper™ ID-X quality flags, of > 95% was obtained. This method involves the lysis of reference samples (tips of cotton swabs, double 4.8 mm diameter Bode or FTA® buccal punches, and single 4.8 mm diameter blood punches on Whatman® 903 or FTA® cards) in 100 µl of Prep-n-Go™ lysis buffer, and amplification of 2 µl of the lysate with 25 cycles. Data collection is optimized by use of the spectral pull-up reduction and off-scale recovery features of the AB3500 Genetic Analyzer Data Collection version 4 software, and by injection of PCR products for two different lengths of time (15 and 7 s). Data was analyzed with GeneMapper™ ID-X version 1.6, which allowed an increased signal dynamic range (maximum peak height of 64000 RFU), and by employing a 10% global filter, and a 20% global stutter filter. Complete GlobalFiler™ profiles were generated from as little as 5 µl of blood, 1 buccal swipe on cotton swab and Bode collectors, and collection of buccal cells deposited on the foam heads of EasiCollect™ FTA® collectors for 5 s. |
| first_indexed | 2024-04-13T17:34:38Z |
| format | Article |
| id | doaj.art-df212705b0fe49c7906e8a701bd45579 |
| institution | Directory Open Access Journal |
| issn | 2665-9107 |
| language | English |
| last_indexed | 2024-04-13T17:34:38Z |
| publishDate | 2022-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Forensic Science International: Reports |
| spelling | doaj.art-df212705b0fe49c7906e8a701bd455792022-12-22T02:37:26ZengElsevierForensic Science International: Reports2665-91072022-07-015100260A single direct amplification method for forensic casework references on a variety of substratesMavis Date Chong0Jeanette Wallin1Corresponding author.; State of California, Department of Justice DNA, Jan Bashinski DNA Laboratory, 1001 W. Cutting Blvd, Richmond, CA 94804, USAState of California, Department of Justice DNA, Jan Bashinski DNA Laboratory, 1001 W. Cutting Blvd, Richmond, CA 94804, USAA single direct amplification method for buccal and blood reference samples, on different substrates, containing a wide range of cellular material, was developed. This method uses the GlobalFiler™ STR kit, which was designed for amplification of previously purified and quantified DNA. Buccal samples on cotton swabs, Bode collectors, and Whatman® FTA® cards, and liquid blood, blood on Whatman® 903 and FTA® cards, and autopsy bloods were examined. A first pass success rate, that is, complete concordant profiles with no GeneMapper™ ID-X quality flags, of > 95% was obtained. This method involves the lysis of reference samples (tips of cotton swabs, double 4.8 mm diameter Bode or FTA® buccal punches, and single 4.8 mm diameter blood punches on Whatman® 903 or FTA® cards) in 100 µl of Prep-n-Go™ lysis buffer, and amplification of 2 µl of the lysate with 25 cycles. Data collection is optimized by use of the spectral pull-up reduction and off-scale recovery features of the AB3500 Genetic Analyzer Data Collection version 4 software, and by injection of PCR products for two different lengths of time (15 and 7 s). Data was analyzed with GeneMapper™ ID-X version 1.6, which allowed an increased signal dynamic range (maximum peak height of 64000 RFU), and by employing a 10% global filter, and a 20% global stutter filter. Complete GlobalFiler™ profiles were generated from as little as 5 µl of blood, 1 buccal swipe on cotton swab and Bode collectors, and collection of buccal cells deposited on the foam heads of EasiCollect™ FTA® collectors for 5 s.http://www.sciencedirect.com/science/article/pii/S2665910722000068Direct GlobalFiler™ amplificationCase reference samplesAB3500 Data Collection version 4GeneMapper ID-X v1.6 off-scale recovery |
| spellingShingle | Mavis Date Chong Jeanette Wallin A single direct amplification method for forensic casework references on a variety of substrates Forensic Science International: Reports Direct GlobalFiler™ amplification Case reference samples AB3500 Data Collection version 4 GeneMapper ID-X v1.6 off-scale recovery |
| title | A single direct amplification method for forensic casework references on a variety of substrates |
| title_full | A single direct amplification method for forensic casework references on a variety of substrates |
| title_fullStr | A single direct amplification method for forensic casework references on a variety of substrates |
| title_full_unstemmed | A single direct amplification method for forensic casework references on a variety of substrates |
| title_short | A single direct amplification method for forensic casework references on a variety of substrates |
| title_sort | single direct amplification method for forensic casework references on a variety of substrates |
| topic | Direct GlobalFiler™ amplification Case reference samples AB3500 Data Collection version 4 GeneMapper ID-X v1.6 off-scale recovery |
| url | http://www.sciencedirect.com/science/article/pii/S2665910722000068 |
| work_keys_str_mv | AT mavisdatechong asingledirectamplificationmethodforforensiccaseworkreferencesonavarietyofsubstrates AT jeanettewallin asingledirectamplificationmethodforforensiccaseworkreferencesonavarietyofsubstrates AT mavisdatechong singledirectamplificationmethodforforensiccaseworkreferencesonavarietyofsubstrates AT jeanettewallin singledirectamplificationmethodforforensiccaseworkreferencesonavarietyofsubstrates |