从法医检材和人类身份鉴定样本中自动纯化DNA,每次可处理1–6个样本或1–14个样本

  • 痕量检材,更高得率
  • 更高信噪比
  • Tip dance方式处理固体样本
  • 可选择使用TE洗脱缓冲液及载体RNA
EZ1 DNA Investigator Kit可从用于人类身份鉴定样本中自动纯化基因组DNA。该试剂盒可配合EZ1 Advanced XL(每批处理1–14个样本)、EZ1 Advanced(每批处理1–6个样本)和BioRobot EZ1(每批处理1–6个样本)工作站使用。纯化过程快速高效,产物DNA在各种下游分析中表现出色,例如定量PCR和STR分析,结果信噪比高。
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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
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.
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Easy and efficient processing of solid materials with the “tip dance” protocol.|Higher yields of concentrated DNA with the large-volume protocol.|EZ1 DNA Investigator trace and “tip dance” protocols.|Improved performance in STR analysis.|
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.|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.|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.|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.)|
Performance
更高的纯化效率,用于敏感的下游分析

EZ1 DNA Investigator Kit含有优化的试剂,可自动纯化获得高产量DNA。在STR分析中获得更高的峰值,提高检测灵敏度(参见"Improved performance in STR analysis")。具有更高的信噪比,确保下游应用中的高灵敏性和优越表现。比较STR分析中的峰高,潜在的DNA的回收率为79%(未显示)。使用TE缓冲液洗脱,结果相似,而且长时间贮存时有更高的稳定性。任何样本的处理过程无遗失(检查任意8个等位基因都有<50的相对荧光单位)。

不同实验方案STR的相对峰高值
实验 试剂盒和卡 方案 洗脱液 相对峰高值 遗漏数
1    EZ1 DNA Tissue Kit和EZ1 DNA Forensic Card 痕量 水  100% 0/8
1 EZ1 DNA Investigator Kit和Card 痕量 176% 0/8
2 EZ1 DNA Investigator Kit和Card 痕量 100% 0/8
2 EZ1 DNA Investigator Kit和Card 痕量 TE 116% 0/8
3 EZ1 DNA Investigator Kit和Card(200 µl) 痕量 100% 0/8
3 EZ1 DNA Investigator Kit和Card(500 µl) 大体积 203%(起始样本量的81% ) 0/8
AmpFlSTR处理的DNA由Buffer G2溶解,最终浓度为5 pg/µl,200 µl(或500 µl的大体积样本)使用特定的试剂盒和方案处理,洗脱体积为50 µl。每个实验中8个峰的峰高都已标准化。(数据由B. Bayer and K. Anslinger,Institute of Legal Medicine,Ludwig Maximilian University, Munich,Germany提供)
tip dance方式处理固体样本,操作简单

除了标准的微量方案,还可选用全自动tip dance实验方案,移液过程中滤嘴相对工作平台可来回移动。这样可在同一管中直接处理固体样本,如:拭子、组织、血斑或烟头。无需事先离心去除会阻塞滤嘴的固态物质(参见。tip dance实验方案简化了操作步骤,在某些情况下能更有效的提取样本提高DNA的产量(参见"Easy and efficient processing of solid materials with the "tip dance" protocol")。

经优化的实验方案,更大的样本容量

大体积实验方案可全自动处理的起始样本量多达500 µl。这不仅可以从含有低浓度DNA的稀释样本,如扩散的污渍中高效纯化DNA,而且能够纯化需要用大体积进行彻底裂解的样本。可处理大容量样本(与标准的微量方案有相同的洗脱体积),就可获得更高产量、更高浓度的DNA,在下游应用中有更高的灵敏度(参见"Higher yields of concentrated DNA with the large-volume protocol")。

有效纯化少量DNA

EZ1 DNA Investigator Kit使用载体RNA,用于提高效率。载体RNA可促进DNA结合到磁珠表面的硅胶上,尤其是含有少量DNA(<100 ng)的样本。公布数据表明载体RNA能够更有效的纯化法医检材样本中的少量DNA,而且对于相同的样本类型可提高DNA产量。载体RNA用于样本裂解也不会影响下游STR分析。

简单的实验设置和操作

使用预设的EZ1 Advanced XL DNA Investigator Card(用于EZ1 Advanced XL)、EZ1 Advanced DNA Investigator Card(用于EZ1 Advanced)和EZ1 DNA Investigator Card(用于BioRobot EZ1)设定纯化方案,十分便利。EZ1 DNA Investigator Kit采用预封装试剂条,减少人工操作的错误和避免缓冲液的污染。通过消除试剂准备和工作站设置时人工操作的失误,以及一个无交叉污染的液体处理,EZ1 Advanced XL、EZ1 Advanced和BioRobot EZ1可确保高度可靠性和标准化。

保证质量

依照QIAGEN's ISO-certified Quality Management System,每批EZ1 DNA Investigator Kit都经过验证,确保始终如一的产品质量。

Principle

该试剂盒配合EZ1 Advanced XL、EZ1 Advanced和BioRobot EZ1工作站,采用硅胶膜磁珠技术全自动纯化基因组DNA。磁珠技术无需重复的离心操作,每批可纯化1–14个样本(EZ1 Advanced XL)或1–6个样本(EZ1 Advanced和BioRobot EZ1)。预封装的试剂条和仪器的自动感控锁可降低污染的风险。

Procedure

将预处理样本加载到工作站,在全自动EZ1 DNA纯化操作后,DNA被一步纯化。洗脱得到的基因组DNA可直接用于各种下游应用。

Applications
使用EZ1 DNA Investigator Kit获得的高品质DNA适用于人类身份鉴定,包括:法医检材、血缘关系鉴定、亲子鉴定和受难者身份鉴定,适用如下技术:
  • STR和SNP基因分型
  • DNA定量检测
  • 线粒体或染色体DNA测序
Feature
Specifications
Applications PCR, real-time RT-PCR, genotyping
Elution volume 50–200 µl
For automated processing BioRobot EZ1
Format Sample tube
Main sample type Forensic samples
Processing Automated
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein DNA
Sample amount <500 µl
Technology Magnetic particles
Yield 150–250 ng

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Validation Reports
2
For the EZ1 DNA Investigator Kit
  • BioRobot EZ1
  • EZ1 Advanced
  • EZ1 Advanced XL

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User-Developed Protocols
3
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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Kit Handbooks
1
For automated purification of DNA from forensic and biosecurity samples using EZ1 instruments
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Supplementary Protocols
4
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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