Quantifiler human dna quantification kit user s manual. Manuals, installation and maintenance downloads for humanscale. Quantifiler human dna quantification kits pcr amplification. This realtime pcrbased kit is designed for the quantification of amplifiable human dna.
Quantifiler human and y human male dna quantification kits. The samples were amplifyed using verity pcr system applyed biosystems. Effects of humic acid on dna quantification with quantifiler. It belongs to human identification solutions category. Wt plus reagent kit user manual pdf, 2 mb publications. Jun 12, 2007 we used quantifilertm human dna quantification kit applied biosystems. Expression wash, stain and scan user manual for cartridge arrays pdf, 2. For detailed information on the separator and column type recommended for a specific reagent, please refer to the data sheet provided with the particular macs cell separation reagent. L of dna extract, quantifiler human primer mix, and quantifiler pcr reaction mix, with thermal cycling conditions according to the manufacturers protocols. Part of the work was presented at the 59th annual meeting of the american academy of forensic sciences, in san antonio, tx, february 1924, 2007, and at the 18th international symposium on human identification, hollywood, ca, october 2007. The quantifiler kits user s manual provides information about and instructions for using the quantifiler human dna quantification kit and the quantifiler y human male dna quantification kit. Implementation of a robotized realtime pcr setup for the.
Each alert wordimportant, caution, warning, dangerimplies a particular level of observation or action, as defined below. Quantifiler human dna quantification kit and quantifiler y human male dna quantification kit user s manual, part number 4344790 rev a. All the samples were resequenced to assess the genotype at each locus tested. The hid evolution system for automation of dna quantification. The quantifiler kits users manual provides information about and instructions for using the quantifiler human dna quantification kit and the quantifiler y human male dna quantification kit. Check the list of manual separators below and choose the one that fits the column type, cell number, sample number, and sample size you are working with. Research day 2011 andrew sawin a comparative analysis of. Cd3 microbeads, human t cells microbeads and isolation.
Dna technical leader quantifiler trio dna quantification kit. Standards should be stored from 25c and expire after one week. Quantifiler human dna quantification kit quantifiler y human. The quantifiler trio dna quantification kit is a robust kit that enables forensic laboratories to simultaneously obtain a quantitative and qualitative assessment of total human and human male dna in a single, highly sensitive realtime pcr reaction. Decisionsupport to help streamline sexual assault sample processing. A total of 75 and 40 standard curves were run for quantifiler human dna quantification kit and quantifiler y human male dna quantification kit, respectively, on different times and days. Automated workstation and abi quantifiler human dna quantification kit the janus automated workstation is a flexible automated liquid handling platform ideally suited for real time quantitative pcr qpcr applications using either total human or yspecific abi quantifiler kits. Optimization of dna concentration to amplify short tandem. Instrument operating software including the qiagility setup manager gene expression, pathogen detection, sample pooling and generic pcr plugin. Internal validation guide of the amp f str identifiler pcr by. All humanscale ergonomic chairs are designed to use the sitters own body weight and the laws of physics to encourage movement and provide unparalleled comfort.
Quantifiler human and y human male dna quantification kits user guide. Text conventions this guide uses the following conventions. The quantifiler human dna quantification kit and the quantifiler y human male dna quantification kit are designed to quantify the total amount of amplifiable human and higher primate dna or human male dna in a sample. Some of the data is also presented in the quantifiler duo dna quantification kit user manual. Target marker size dye human dna rpph1 ribonuclease p rna component h1 140 bp vic human male dna sry sex determining region y bp fam ipc artificial template bp ned. Quantifiler hp dna quantification kit 400 reactions 4482911 quantifiler trio dna quantification kit 400 reactions 4482910 hid 7500 calibration starter kit na a25019 hid realtime pcr analysis software v1. Included are protocols for setting up the abi prism 7000 and 7900ht sequence detection systems.
Quantifiler duo dna quantification kit user guide chapter 1 overview realtime data analysis 1 the 7500 system uses the data obtained from the pure. Research day 2011 andrew sawin a comparative analysis. Human dna quantification kit and quantifiler y human male dna quantification kit, users manual, rev. Quantifiler kits user s manual ix safety safety alert words four safety alert words appear in applied biosystems user documentation at points in the document where you need to be aware of relevant hazards. Apr 14, 2009 initially, a humanspecific realtime pcr assay, a quantifiler human dna quantification kit applied biosystems, foster city, ca, was used to quantify the genomic dna in 100 selected samples. Quantifiler hp and quantifiler trio dna quantification kits. Dna extracted from formalinfixed tissues may also provide an alternative source of genetic material for medical diagnosis and forensic casework, such as identifying unknown embalmed human remains.
Initially, a human specific realtime pcr assay, a quantifiler human dna quantification kit applied biosystems, foster city, ca, was used to quantify the genomic dna in 100 selected samples. A guiding tool for short tandem repeat genotyping of forensic samples. Evaluation of four automated protocols for extraction of dna. Manual cell separators for fast and gentle cell isolation. Exogenous male dna was quantified using a quantifiler y human male dna quantification kit applied biosystems, foster city, ca. Eight standards were created via serial dilution of the quantifiler human dna standard according to the quantifiler kit user manual. Duo dna standard and quantifiler duo dilution mix in the reaction use 10. For the quantifiler trio kit only, the relative quantities of human male and. Since automation of pcrsetups reduces the manual workload and the risk for human errors, we have chosen to validate the pcrsetup of the quantifiler trio kit on a small benchtop liquid handler, the biomek 4000 beckman coulter. Refer to the quantifiler kits users manual or the appropriate instrument quick reference card for instructions on setting up the plate document, running. Effects of humic acid on dna quantification with quantifiler human dna quantification kit and short tandem repeat amplification efficiency. Incorrect dna quantification due to the presence of pcr inhibitors may affect experiment results. Validation of automated pcrsetup of the quantifiler trio. Therefore automation of quantitative realtime pcr setup was an important component of our dnaanalysis automation concept.
International journal of legal medicine ausgabe 62012 autoren. Our broad portfolio of available kits spans multiple therapeutic areas including cardiology, infectious disease, inflammation, neurology, and oncology. View online or download applied biosystems 7500 fast installation and maintenance manual, getting started. Workflow materials figure 1 the quantifiler human dna quantification kit and quantifiler y human male.
This standard is generated from pooled human male genomic dna. Information regarding preparation of standards should be recorded in quantifiler duo dna quantification standards log. The human dnaspecific assay includes primers specific for a particular human or higher primate dna sequence and a taqman probe that binds to that sequence. Quantifiler human dna quantification kit 400 x 25 l 4343895 quantifiler y human male dna 400 x 25 l 4343906 quantification kit quantifiler human dna kit users manual 4344790 abi prism 7000 sequence detection system laptop 43492 abi prism 7000 sequence detection system tower 4349117. Quantifiler human dna quantification kit applied biosystems as a screening kit for dna.
Part number 4391294 rev 042008 part number 4391294 rev 042008 barbisin, maura, et al. Quantifiler human dna quantification kit genetics supplement. Applied biosystems, foster city applied biosystems 2005 quantifiler human dna quantification kit. Whole genome amplification and realtime pcr in forensic. The quantifiler duo dna quantification kit users manual provides information about and instructions for using the quantifiler duo dna quantification kit. Refer to the quantifiler kits user s manual or the quantifiler human dna quantification kits pcr amplification. Human dna quantification kit and the quantifilerz y human male dna quantification kit used with the abi prism. Here we show the implementation of a robotized setup for the quantifiler human dna quantification kit. Thermo fisher scientific provides wide range of products under applied biosystem brand which includes quantifiler duo dna quantification kit user. The quantifiler duo kit will detect samples containing a dna concentration of less than 23pgl. Select manual baseline and enter 3 to 15 as cycle range.
All samples were quantified using a quantifiler human dna quantification kit applied biosystems according with manufacturers protocols. Graphic from the quantifiler kitsquantifiler human dna quantification kit and quantifiler y human male dna quantification kit. Rf exposure this equipment complies with fcc radio frequency exposure limits set forth for uncontrolled environment, when installed and operated so that a minimum distance of 20 cm is kept towards the nearest human body. Quantifiler human dna quantification kit quantifiler, quantifiler y. Quantifiler observations of relevance to forensic casework. The quantifiler human dna quantification kit and the quantifiler y human male.
Hp and trio kits enable efficient and accurate quantification of human dna and are the first kits to provide a quality index to detect the presence of degraded dna along with pcr inhibitors. To obtain optimal results it is critical that the procedures detailed in the user s manual are followed. Walkaway qpcr setup using the janus automated workstation. Budowle b, moretti tr, keys km, koons bw, smerick jb. The reproducibility of the quantifiler duo dna quantification kit on the 7500 realtime pcr instrument will be further evaluated with mock case samples and various dilutions of single. See quantifiler duo dna quantification kit users manual for information regarding the procedure. The range of slope and r 2 values observed are summarized in table 2. Effects of humic acid on dna quantification with quantifiler human dna quantification kit and short tandem repeat amplification efficiency zeitschrift. Provides complimentary total human and male dna quantification. The effects of metal ion pcr inhibitors on results obtained. Developmental validation of the quantifiler duo dna quantification kit for simultaneous quantification of total human and human male dna and detection of pcr inhibitors in biological samples. Audience this manual is intended for scientists who use the quantifiler duo dna quantification kit for the quantification of human dna extracted from a variety of sample types. Quantifiler duo dna quantification kit using the ab 7500. The primary quantification targets small autosomal and y consist of relatively short amplicons 75 to 80 bases to improve the detection of degraded dna.
The master mixes were prepared using the qiagility and two qas run files created during the validation. Sureselect human all exon v7 agilents latest exome, the sureselect human all exon v7, is a comprehensive exome that focuses on interpretable part of the genome, and also provides a costeffective hybridcapture solution. The effects of metal ion pcr inhibitors on results. This realtime pcrbased kit is designed for the quantification of amplifiable human dna that is present in a sample, and has been validated for human identification applications. Quantifiler human dna quantification kit quantifiler y.
Quantifiler human dna quantification kits on the abi prism 7000. Y human male dna quantification kit users manual, part number 4344790 rev a. List of components the smarter human tcr ab profiling kit consists of the smarter human tcr ab profiling kit components not. The qiagility was programmed to prepare the standards following the dilution scheme given in the quantifiler kit users manual. The effects of metal ion pcr inhibitors on results obtained with the quantifiler.
Quantifiler human dna quantification kits on the abi prism. Applied biosystems 7500 realtime pcr system forensic. Quantifiler human dna standard both quantifiler kits utilize the same reagents and allow the user to select the appropriate quantifiler primer mix, which contains a preformulated mixture of primers and probes that amplify either the human telomerase reverse transcriptase gene quantifiler human kit or the sry gene quantifiler y kit. This study used the quantifiler duo dna standard to assess reproducibility. The run files allow the user to define the number and type of samples needed for each plate. Seung bum seo, hye young lee, ai hua zhang, hye yeon kim, dong hoon shin, soong deok lee. Formalin fixation is commonly used to preserve tissue sections for pathological testing and embalming cadavers for medical dissection or burial. Quantifiler kits quantifiler human dna quantification kit and quantifiler y human male dna quantification kit users manual. Quantifiler human dna quantification kit thermo fisher scientific. The users manual for human body chinese edition qingzhong wu on. Graphic from the quantifiler kits quantifiler human dna quantification kit and quantifiler y human male dna quantification kit. The ipc assay includes a unique synthetic dna sequence as well as primers and a taqman.
Quantifiler duo dna quantification kit users manual. Validation studies of the ctt str multiplex system. Quantifiler duo dna quantification kit thermo fisher. Duplication of any part of this document is permitted for classroom use only. Validated on the 7500 real time pcr system using sds v1. Quantifiler hp and trio dna quantification kits user guide.
Collecting and analyzing dna evidence from fingernails. The human kit assay is intended for general use as a preliminary. The quantifiler duo dna quantification kit enables forensic laboratories to simultaneously obtain a quantitative and qualitative assessment of total human and human male dna in a single, highly sensitive realtime pcr reaction. Modifications not expressly approved by verizon telematics inc. Configuration of quantifiler duo kit similar amplicon sizes to avoid preferential pcr. To the instructor immunology is the study of the immune system and how the body protects itself against. According to manufacturers instructions quantifiler kit assays.
Quantifiler duo dna quantification kit user guide pub. Manual processing for the setup of quantification reactions can be time consuming and labor intensive. Smarter human tcr ab profiling kit user manual 012618 takara bio usa, inc. This validation introduces the newly released applied biosystems 7500 realtime pcr system with v1. This quick reference card covers setting up the plate document, running pcr, and viewing result s.
For quantification purposes absolute quantitation method is used, realtime quantitative pcr assay with a fluorogenic taqman probe targeting the human telomerase reverse transcriptase gene htert and supported by quantifiler human dna quantification kit on instrument abi prism 7000 sequence detection system applied bioystemsab, pn 4330087 with sds software v1. The quantifiler duo dna quantification kit user s manual provides information about and instructions for using the quantifiler duo dna quantification kit. Internal validation guide of the amp f str identifiler pcr. A master mix, consisting of quantifiler pcr reaction mix and quantifiler human primer mix applied biosystems, was prepared according to the manufacturers instructions and distributed into a 96well optical reaction plate. The dynamic range of the quantifiler duo assay is 23pg50ngl and is the same as for the existing quantifiler kits. Developmental validation of the quantifiler duo dna. Amplification was performed using the quantifiler kit. The human dna standard in the quantifiler human and quantifiler y kits was used to prepare the serial dilutions to serve as known values. The quantification assay was performed in a total volume of 25. Validation information to aid forensic dna laboratories nist. Implementation of a robotized realtime pcr setup for the use.
1056 945 1329 1486 787 1064 573 1315 464 149 1120 1510 648 1040 671 233 1280 885 1015 1240 514 829 414 279 579 682 882 1146 852 1372 1286 1514 1217 208 902 411 1113 1120 468 1020 529 1038 1428 197