Julianne Henry, Lenara Scandrett, Assessment of the Yfiler® Plus PCR amplification kit for the detection of male DNA in semen-negative sexual assault cases, Science & Justice, 10. 64% Yfiler average ~0. Yfiler® vs Y23 “Premier Public Safety Services” 5 • Dynamic range and sensitivity – Both kits appeared to have roughly equal sensitivity and dye balance. • Additional highly polymorphic loci: DYS437, DYS448, DYS456, DYS458, DYS635 (Y GATA C4), and Y GATA H4. Approximately 50 injections of the Yfiler TM ladder and 20 injections of a sample amplified using the Yfiler TM kit were run on four 310 Genetic Analyzers. Appendix Q Quality Control Check of Yfiler Plus Kit. · For standard reactions, volumes and amplification conditions as defined in the AmpFℓSTR ® Yfiler™ PCR Amplification Kit User’s Manual were followed. 0ng of male DNA (Yfiler® Plus PCR Amplification Kit) in the presence of increasing amounts of common inhibitors.
Key features of the Yfiler Plus PCR Amplification Kit:. This paper describes our validation results of the Yfiler ® Plus kit for forensic casework analysis. The GlobalFiler Amplification kit is used for Autosomal (STR) DNA analysis, and the Yfiler Plus Amplification Kit is used for male-specific (Y-STR) DNA analysis. Profiler™ Plus and COfiler™. Forty microliters of buccal pool were deposited on NUCLEIC-CARD™ COLOR, while 40ul of each blood sample were deposited on NUCLEIC-CARD™ White cards for each donor. – Both kits started exhibiting dropout at approximately 0. Product description The Yfiler® Kit contains all the necessary reagents for the amplification of human male-specific genomic DNA. Yfiler ® Plus PCR Amplification Kit using a new 6-dye system detects 27 Y-STR loci.
Table 3 - Yfiler™ PR amplification reagents for one sample Reagent Per reaction Yfiler™ P R Reaction Mix 9. YFiler Plus PCR Amplification using GeneAmp PCR System 9700 Applied Biosystems 3500 Genetic Analyzer Procedures using GlobalFiler and Yfiler Plus GeneMapper ID-X Software Protocol for the Analysis of GlobalFiler and Yfiler Plus STR Data. DNA_5 PCR – Yfiler Plus DNA_6 3500 GA – GlobalFiler and YfilerPlus DNA_7 GMID-X Analysis Casework Standard Operating Procedures Section Number Section Name CASE_1 Notes CASE_2 Photography CASE_3 Serology CASE_4 DNA Analysis CASE_5 DNA Interpretation CASE_6 DNA Statistics CASE_7 Report Wording CASE_8 Retention of Samples. Adventitious match of Y-Filer profiles between two ethnic groups. 05 uL Semen (10-20 pg male DNA added to STR reaction) • On average, 2 loci from male donor were recovered. PowerPlex™16 HS at 30 cycles and Identifiler™ Plus at 29 seem optimal. 05ng of input DNA.
5ng appeared to be ideal target template for single source samples. He explained, "In the time since my affidavit, some. · 15 Proprietary & Confidential Results: YFiler Plus, Samples with 0. Applied Genetics Rapidly Mutating (RM) Y-STRs. Until YFiler Plus-specific θ estimates are available, USYSTR Database users who submit a 27-locus YFiler Plus haplotype can still obtain a count of the number of times the haplotype has been observed in the database, its relative frequency and the variation in this estimate based upon database size. · either the "PowerPlex Y23" or the "Yfiler Plus," that had become available in but that, under national accreditation standards, required a process of internal validation by a given laboratory before it could adopt the test for its own use. For manual operation, adding a role letter is a way to add a suspect or reference allele.
In this comparative study, Y-STR typing with both Yfiler™ and Yfiler™ Plus (Thermo Fisher Scientific, Waltham, MA, USA) was performed on a variety of human skeletal remains, including samples from the American Civil War (1861–1865), the late nineteenth century gold rush era in Deadwood, SD, USA (1874–1877), the Seven Years’ War (1756. • Blood samples collected on untreated paper substrates and treated with Prep-n-Go™ Buffer. The Yfiler(®) Plus PCR Amplification Kit is a dual application assay designed to amplify DNA from extracted casework and database samples from storage cards and swab lysates via direct. Table of Contents. · One primer for TPOX and DYS391 has also been redesigned from Identifiler™ Plus and Yfiler™ Kits, respectively, to increase the size of the amplicon and facilitate optimal marker spacing 14, 17.
NDIS Approved Expert System. and 25 loci in the Yfiler® Plus Kit. Manual of Style, 10th edition,. These highly variable regions among individuals make them effective for human identification purposes. y filer plus manual AmpFLSTR™ Yfiler™ PCR Amplification Kit 2. (Promega) and the YFiler Plus (Thermo Fisher) chemistries. The Yfiler™ Plus PCR Amplification Kit is optimized to allow direct amplification from the following types of single-source samples without the need for sample purification: • Blood and buccal samples on treated paper substrates. This QA Manual has been approved by the DNA Casework Supervisor, CODIS Administration, DNA Technical Leader, Laboratory Quality Assurance Manager, Assistant Director, and Executive Director and is accepted as routine operating policy of the Forensic DNA Section within the Arkansas State Crime Laboratory.
The Yfiler ® Plus PCR Amplification Kit is a dual application assay designed to amplify DNA from extracted casework and database samples from storage cards and swab lysates via direct amplification. Read the Safety Data Sheets (SDSs) and follow the handling instructions. The 5 dyes used in the GlobalFiler™ Kit to label amplified sample products are 6-FAM™, VIC™, NED™, TAZ™, y filer plus manual and SID™. 2kV, 24-second injection). Yfiler Plus Point of Comparison: DYS458 has highest mutation rate in Yfiler at 0. 37 — mostly bugfix YFiler Plus 37. While PowerPlex™16 HS exhibited better sensitivity than Identifiler™ Plus at its 32 cycles parameter, that better sensitivity comes with much a higher amount of artifacts. Metapopulation &92; Dataset Minimal PowerPlex Y Yfiler PowerPlex Y23 Yfiler Plus Maximal; Worldwide C, Aug-C, K, Avg-DL.
Y-STR panel (European minimal haplotype plus DYS438 and DYS439). Used With: AmpFLSTR™ Yfiler™ PCR Amplification Kit 2. DNA Profiles : CODIS, Robotics, Genetic Analyzer, Thermal Cyclers, Globalfiler Express Amp Kit; Yfiler Plus Amp Kit; Data Interpretation System. GeneMarker® HID human identity software is an excellent choice for all forensic profiling applications. The Yfiler Plus Kit shows very high concordance to the Yfiler Kit but discordance with the PowerPlex(®) Y23 Kit at the DYS481 locus was observed in 2 out of 30 samples tested. The AmpFlSTR®™Yfiler PCR Amplification Kit User’s Manual provides information about the Applied Biosystems instruments, chemistries, and software associated with the AmpF lSTR Yfiler PCR Amplification Kit (Yfiler kit).
Compared to the Yfiler PCR Amplification Kit, the new multiplex shows increased discrimination of male lineages and also improved performance in inhibited samples, improved balance in male DNA samples mixed with female DNA at ratios >1:1000, and faster time to results. This developmental validation work follows the SWGDAM guidelines and demonstrates that the assay is robust and suitable for use on forensic casework and database samples. StepOnePlus™ Real-Time PCR System Spectral Calibration Kit (96 well) Catalog NumberPub.
29 6 automated globalfiler and y-filer plus testing of. Only one extract contained sufficient DNA for it to be amplified at the optimal 400 pg DNA template amount (the optimal input DNA was determined by in-house validation of the Yfiler and. AmpFLSTR™ SGM Plus™ PCR Amplification Kit User&39;s Manual 2. Amplification of 500pg of male DNA (PowerPlex® Y23 System) or 1.
Information on the National Database "China" The National Database "China" consists of 115703 haplotypes (at least minimal) and 182 unique population samples. DNA 10-18 YSTR DNA Amplification Using Yfiler Plus DNA 11-01 CODIS Casework--Administration DNA 11-02 CODIS Casework - Analysts DNA 11-03 y filer plus manual CODIS Familial Searching DNA 12-01 Proficiency Testing DNA 12-02 Training DNA 12-03 Report Writing and Reviews DNA 12-04 Subcontracting and Outsourcing DNA 13-01 STRMix Interpretation. The amount of reagents for one amplification reaction is listed in Table 3. Consult the amplification documentation for the exact amount of Yfiler™ primers, reaction mix and ABI Taq Gold to add.
We examined sizing precision, baseline noise, sensitivity, stutter ratios, peak height ratios, spectral pull-up, the tolerance to PCR inhibitors, ability to obtain results from aged bloodstains and mixed DNA samples, and artifact peaks. manual/automated extraction: - Promega SwabSolution - Prep-N-Go - QIAmp DNA Investigator (nail only) Documented in-house methods SOP ref: PRC521, PRC 545 using manual/automated amplification and the following chemistry: - Fusion STR Multiplex - Verifiler Express - HDPlex - Yfiler Plus Documented in-house methods SOP ref:. globalfiler and y-filer plus of evidence samples. · Profiling using the Yfiler Plus kit occurred up to 3 years later than profiling in Yfiler due to the later release and implementation of Yfiler Plus at FSSA. DNA was amplified for 30 cycles, and amplified products were separated on an Applied Biosystems® 3500xL Genetic Analyzer (1.
Would not recommend 32 cycle protocol in casework. The modified reactions maintained the 25-μL total reaction volume of the standard protocol but the recommended Taq concentration was doubled and thus a maximum volume of 9. AmpFLSTR™ Yfiler™ PCR Amplification Kit User&39;s Manual 2. Details on National Database . 2mm discs/card were punched with the. The Yfiler Plus PCR Amplification Kit is a 6-dye multiplex assay for short tandem repeats (STRs) that allows amplification from multiple male-specific sample types, such as male–male and male–female mixtures, and direct PCR amplification from single-source samples. dna methods manual.
2 µL Yfiler™ Primer Set 5. The OSBI FBU utilizes two amplification kits from Applied Biosystems. y filer plus manual Y Filer Amp Kit, Genetic Analyzer, Thermal Cyclers, Data Interpretation System Individual Characteristics Database : CODIS Methods Manual. The Applied Biosystems™ Yfiler™ Plus PCR Amplification Kit is a 6-dye, short tandem repeat (STR) multiplex assay optimized to allow amplification from multiple male- specific sample types such as male-male, male-female mixtures. Text ConventionsThis guide uses the following conventions: •Bold indicates user action.
The average, standard deviation, maximum, minimum, and range of the size in base pairs were determined for the largest and smallest allele at each locus in. Forensic Biology Quality Assurance Manual.
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