For automated purification of DNA from 1–6 or 1–14 forensic and human identity samples per run
- More efficient yields from trace casework samples
- Higher signal-to-noise ratios
- “Tip dance” protocol for processing solid material
- Optional TE elution and use of carrier RNA
The EZ1 DNA Investigator Kit reproducibly automates purification of genomic DNA from reference and casework samples in human identity testing. The kit can be used with the EZ1 Advanced XL (1–14 samples per run), EZ1 Advanced (1–6 samples per run), and BioRobot EZ1 (1–6 samples per run) instruments. Purification is fast and efficient, and purified DNA performs well in downstream analyses, such as quantitative PCR and STR analysis, with high signal-to-noise ratios.
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EZ1 DNA Investigator Kit (48)
For 48 preps: Reagent Cartridge (DNA Investigator), Disposable Filter-Tips, Disposable Tip-Holders, Sample Tubes (2 ml), Elution Tubes (1.5 ml), Buffer G2, Proteinase K, Carrier RNA
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952034
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The EZ1 DNA Investigator Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
“Tip Dance” Protocol を用いて簡単で効率的な固体サンプル処理|Large-Volume Protocolを用いて濃縮されたDNAを高収量で精製|EZ1 DNA Investigator traceおよび “tip dance” プロトコール|STR解析における改善された解析結果|
タバコの吸殻からの紙あるいは各サンプルあたり3枚のペーパーディスクそれぞれに50 ng のDNAをスポッティングした。 Proteinase Kによる分解後、サンプルを95℃で5分間インキュベートした。 半分のサンプルから固体サンプルを除去し、EZ1 DNA Investigator Kit とstandard trace protocol を用いて精製した(Trace)。 サンプルの残りの半分を、サンプルチューブから固体サンプルを除去することなく、EZ1 DNA Investigator Kit と“Tip dance”プロトコールを用いて調製した(Tip dance)。 DNA収量をリアルタイム定量PCRで定量した。|最終濃度50 pg/μl になるようにDNAをBuffer G2で希釈し、表記量をstandard trace protocolプロトコール(Trace)、あるいはlarge-volume protocol (LV)とEZ1 DNA Investigator Kit を用いて調製した。 全てのサンプルを50 μl の水で溶出し、そのうち5 μl をリアルタイム定量PCRで定量した。|標準的なtrace プロトコールに加え、ピペッティング中にワークテーブルに対してフィルターチップが前後運動する全自動の“tip dance” プロトコールがオプションとして使用できる。このプロトコールにより、スワブ、繊維、血液ディスク、タバコの吸殻のような固体サンプルをサンプルチューブ内で直接処理することが可能である。チップを詰まらせる可能性のある固体サンプルを除去するために事前に遠心操作を行なう必要はない。|AmpFlSTR コントロールDNA(1 ng) を200 μl のBuffer G2 で希釈し、EZ1 DNA Investigator Kit および trace protocol を用いてEZ1 Advancedワークステーションで精製した。 DNAを50 μl の水で溶出し、10 μl (200 pgのDNAに相当) をSTR解析に用いた。 PCR産物は、ABI PRISM 310 Genetic AnalyzerとGenotyper ソフトウェアとの組み合わせにより解析した(データ提供: B. Bayer and K. Anslinger, Institute of Legal Medicine, Ludwig Maximilian University, Munich, Germany)。|
パフォーマンス
More efficient yields for sensitive downstream analyses
The EZ1 DNA Investigator Kit provides optimized chemistries for automated DNA purification with more efficient yields. STR analysis shows higher peaks for more sensitive detection (see figure "Improved performance in STR analysis", see table "Relative STR peak heights with different protocols"). The higher peaks demonstrate the improved signal-to-noise ratio for highly sensitive detection and high performance in downstream applications. Comparison of peak heights in STR analysis showed recovery of 79% of the potential DNA yield (not shown). Optional elution in TE buffer gives comparable performance (see table "Relative STR peak heights with different protocols") and provides increased stability for long-term storage of small amounts of purified DNA. There were no dropouts (<50 relative fluorescent units for any of the 8 alleles investigated) for any of the samples processed (see table "Relative STR peak heights with different protocols").
| 1 |
EZ1 DNA Tissue Kit and EZ1 DNA Forensic Card |
Trace |
Water |
100% |
0/8 |
| 1 |
EZ1 DNA Investigator Kit and Card |
Trace |
Water |
176% |
0/8 |
| 2 |
EZ1 DNA Investigator Kit and Card |
Trace |
Water |
100% |
0/8 |
| 2 |
EZ1 DNA Investigator Kit and Card |
Trace |
TE |
116% |
0/8 |
| 3 |
EZ1 DNA Investigator Kit and Card (200 µl) |
Trace |
Water |
100% |
0/8 |
| 3 |
EZ1 DNA Investigator Kit and Card (500 µl) |
Large-volume |
Water |
203% (81% normalized to starting volume) |
0/8 |
Easier handling of solid materials with "tip dance" protocol
In addition to the standard trace protocol, an optional, fully automated "tip dance" protocol can be used, where the filter-tip moves back and forth relative to the worktable platform while pipetting. This enables processing of solid materials, such as swabs, fabrics, blood discs, or cigarette butts, directly in the sample tube. There is no need for prior centrifugation to remove solid materials that could clog the tip (see flowchart "EZ1 DNA tissue trace and “tip dance” protocols"). The "tip dance" protocol simplifies handling and, in some cases, increases DNA yield with more efficient extraction of the sample (see figure "Easy and efficient processing of solid materials with the "tip dance" protocol").
Optimized protocol for larger sample volumes
A large-volume protocol enables fully automated processing of starting volumes up to 500 µl. This not only allows efficient DNA purification from dilute samples with low concentrations of DNA, such as diffuse stains, but also enables purification from samples that require larger volumes for thorough lysis. The ability to process larger sample volumes — with the same elution volume as the standard trace protocol — enables higher yields of more concentrated DNA for greater sensitivity in downstream applications (see figure "Higher yields of concentrated DNA with the large-volume protocol", see table "Relative STR peak heights with different protocols").
Efficient purification of low amounts of DNA
The EZ1 DNA Investigator Kit now includes carrier RNA for increased efficiency. Carrier RNA enhances binding of DNA to the silica surface of the magnetic particles, especially if the sample contains low amounts of DNA (<100 ng). Published data suggest that addition of carrier RNA enables more efficient isolation of low amounts of DNA from forensic samples and may, for some sample types, provide improved DNA yields. Addition of carrier RNA to sample lysates did not interfere with downstream STR analysis.
Easy protocol setup and handling
The preprogrammed EZ1 Advanced XL DNA Investigator Card (for use with the EZ1 Advanced XL), EZ1 Advanced DNA Investigator Card (for use with the EZ1 Advanced), and EZ1 DNA Investigator Card (for use with the BioRobot EZ1) make setting up the purification protocols as easy as making a credit card transaction. Prefilled, sealed reagent cartridges in the EZ1 DNA Investigator Kit help to prevent human error during setup of the instrument and prevent contamination of buffers. By eliminating human error in reagent preparation and workstation setup, and by providing cross-contamination-free liquid handling, the EZ1 Advanced XL, EZ1 Advanced, and BioRobot EZ1 provide high levels of reliability and standardization.
Quality guaranteed
In accordance with QIAGEN's ISO-certified Quality Management System, each lot of EZ1 DNA Investigator Kits is tested against predetermined specifications to ensure consistent product quality.
原理
The EZ1 Advanced XL, EZ1 Advanced, and BioRobot EZ1 provide fully automated purification of genomic DNA using silica-coated magnetic particles. Magnetic-particle technology eliminates tedious centrifugation steps, and the easy-to-use instrument allows purification of 1-14 samples (EZ1 Advanced XL) or 1–6 samples (EZ1 Advanced and BioRobot EZ1) in a single run. The automatic sensor-controlled lock of the instrument door and the use of sealed reagent cartridges reduce the chances of environmental contamination.
操作手順
Pretreated samples are loaded onto the instrument, and genomic DNA is purified in one step following the automated EZ1 DNA procedure. Genomic DNA is eluted, ready to use in all downstream applications.
アプリケーション
The high-quality DNA obtained using the EZ1 DNA Investigator Kit is suited for human identity applications including forensics, kinship or paternity testing, and disaster victim identification, and is used in techniques such as:
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STR and SNP genotyping
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DNA quantitation
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Mitochondrial or chromosomal DNA sequencing
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Feature
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Specifications
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Applications
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PCR, real-time RT-PCR, genotyping
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Elution volume
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50–200 µl
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For automated processing
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BioRobot EZ1
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Format
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Sample tube
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Main sample type
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Forensic samples
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Processing
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Automated
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Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein
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DNA
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Sample amount
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<500 µl
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Technology
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Magnetic particles
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Yield
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150–250 ng
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This protocol has been adapted by the San Diego Police Department (SDPD) Crime Laboratory from the standard QIAGEN pretreatment for casework samples, and is intended for the lysis and extraction of DNA from forensic samples of fired (casings) and unfired (cartridges) ammunition.
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This protocol has been adapted by the North Louisiana Criminalistics Laboratory from the pretreatment for hair and is for the lysis and extraction of DNA from forensic samples of hair in less time than the standard EZ1 DNA Investigator protocol pretreatment.
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This protocol has been adapted by the North Louisiana Criminalistics Laboratory from the pretreatment for epithelial cells mixed with sperm cells and is for the lysis and extraction of DNA from a sperm pellet after differential separation.
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This protocol is adapted from the QIAGEN RNeasy MinElute Cleanup protocol and is intended for use as a guideline for the purification of total RNA from waste fractions obtained from a sample first purified using the EZ1 DNA Investigator protocol.
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This protocol has been adapted to allow the user to replace the epithelial transfer and sperm pellet wash steps with an automated protocol using the QIAcube Connect.
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For automated purification of DNA from forensic and biosecurity samples using EZ1 instruments
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Supplementary protocol using the EZ1 Advanced DNA Investigator Large-Scale Bone Card or the EZ1 Advanced XL DNA Investigator Large-Scale Bone Card
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Supplementary protocol using the Flip Cap Rack and the EZ1 Advanced XL Flip Cap Card.
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Supplementary protocol using the Investigator Lyse&Spin Basket Kit and EZ1 Advanced XL
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Supplementary protocol using the EZ1 Advanced or EZ1 Advanced XL with the EZ1 DNA Investigator Kit
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Images
Easy and efficient processing of solid materials with the “tip dance” protocol.
Papers from cigarette butts or 3 paper disks per sample were spotted with 50 ng DNA per sample. After proteinase K digestion, the samples were incubated at 95°C for 5 minutes. Solid materials were removed from half of the samples, which were then processed using the EZ1 DNA Investigator Kit with the standard trace protocol (Trace). The remaining half of the samples was processed using the EZ1 DNA Investigator Kit with the "tip dance" protocol, without removing solid materials from the sample tubes (Tip dance). DNA yields were quantified by real-time, quantitative PCR.
Higher yields of concentrated DNA with the large-volume protocol.
DNA was diluted in Buffer G2 to a final concentration of 50 pg/μl, and the indicated volumes were processed using the EZ1 DNA Investigator Kit with the standard trace protocol (Trace) or the large-volume protocol (LV). All samples were eluted in 50 μl water, and 5 μl was quantified using real-time, quantitative PCR.
EZ1 DNA Investigator trace and “tip dance” protocols.
In addition to the standard trace protocol, an optional, fully automated "tip dance" protocol can be used, where the filter-tip moves back and forth relative to the worktable platform while pipetting. This enables processing of solid materials, such as swabs, fabrics, blood discs, or cigarette butts, directly in the sample tube. There is no need for prior centrifugation to remove solid materials that could clog the tip.
Improved performance in STR analysis.
AmpFlSTR control DNA (1 ng) was diluted in 200 μl Buffer G2 and purified using the EZ1 DNA Investigator Kit and the trace protocol on the EZ1 Advanced workstation. DNA was eluted in 50 μl water, and 10 μl (corresponding to 200 pg DNA) was used for STR analysis. PCR products were analyzed on an ABI PRISM 310 Genetic Analyzer with Genotyper software. (Data kindly provided by B. Bayer and K. Anslinger, Institute of Legal Medicine, Ludwig Maximilian University, Munich, Germany.)
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