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RNeasy 96 BioRobot 9604 Kit

在BioRobot 9604工作站上高通量自动化从细胞中少量制备RNA

  • 在BioRobot 9604工作站上高通量自动纯化RNA
  • 样本处理快速方便
  • 无有机物提取或沉淀
  • 从10至500,000个细胞中纯化获得RNA,产量重复性好
  • 得到的高品质RNA适合各种下游应用
Cat No./ID: 967142
RNeasy 96 BioRobot 9604 Kit (12)
For 12 x 96 total and cytoplasmic RNA preps on the BioRobot 9604 or BioRobot 3000: 12 RNeasy 96 Plates, Elution Microtubes CL, Caps, S-Blocks, AirPore Tape Sheets, RNase-Free Reagents and Buffers
0
High-quality RNA for sensitive analysis of a low-copy transcript.
RNA was isolated from 1 to 1 x 105 HeLa S3 cells using the RNeasy 96 BioRobot 8000 procedure. Total RNA was eluted in 100 µl RNase-free water, and 5 µl was used for RT-PCR. Quantitative, real-time one-step RT-PCR analysis was carried out on an ABI Sequence Detection System using the QuantiTect Probe RT-PCR Kit with primers and probe specific for the low-copy c-fos transcript. [A] Amplification plot. [B] CT values. Error bars represent standard deviation from 8 different samples for each cell number.
1
Reproducible yields of high-quality RNA.
[A] Agarose gel analysis. HeLa cells (5 x 105) from a homogeneous cell culture were pelleted in each well of a 96-well cell-culture plate and the total RNA isolated using the RNeasy 96 system. Half of each eluate was analyzed on a formaldehyde agarose gel. [B] The corresponding northern blot, hybridized with a 32P-labeled GAPDH probe. [C] Analysis of RNA yields. Each square represents the yield from a preparation originating from one well of the 96-well cell-culture plate.
2
RT-PCR of RNA from as few as 100 cells.
RT-PCR of cytoplasmic RNA isolated with the RNeasy 96 Kit from the indicated numbers of HeLa cells. 10 µl (1/10) of the eluate was reverse-transcribed with oligo-dT primer, and 5 µl (1/5) of the cDNA mix was used in a 100 µl PCR in which a 214 bp fragment of beta-actin was amplified. C-: no template; C+: beta-actin in vitro transcript; M: 100 bp DNA ladder.
3
Repeatability of automated RNA purification.
RNA was isolated from 96 aliquots (5 x 104 cells each) of a HeLa S3 cell culture using the RNeasy 96 system and the RNeasy 96 BioRobot 8000 procedure. Quantitative, real-time one-step RT-PCR was set up in the same protocol on the BioRobot 8000 workstation, using the QuantiTect Probe RT-PCR Kit with primers and dual-labeled probe specific for the low-copy c-myc transcript. Threshold cycles (CT) are shown for all 96 samples. The mean CT was 21.34 ± 0.34 (mean ± standard deviation), representing a CV of 1.6%.
4
RNeasy 96-well plate.
Performance
RNeasy 96系列试剂盒可标准化、高通量的从样本中可靠纯化高品质RNA,适用于高通量基因表达分析。纯化得到的RNA适用于real-time RT-PCR和微芯片分析等敏感的下游应用(参见"High-quality RNA for sensitive analysis of a low-copy transcript")。样本量覆盖10到5 x 105个细胞(参见"RT-PCR of RNA from as few as 100 cells"),可从样本中纯化得到高品质RNA(参见"Reproducible yields of high-quality RNA")。在整个纯化过程中,个体差异水平低;使用QuantiTect Probe RT-PCR Kit在纯化过程结束时可轻松的达到TaqMan循环阈值而变异系数低于3%(参见"Repeatability of automated RNA purification")。
Principle

RNeasy 96系列试剂盒可标准化、高通量的从样本中可靠纯化高品质RNA(参见下表)。RNeasy 96纯化产品的纯化速度和效率的关键是简洁的流程。可在96孔板细胞培养皿中培养细胞并直接裂解。将裂解物转移至RNeasy 96孔板中后(参见"RNeasy 96-well plate"),每个孔中的RNA与硅胶模结合,污染物被洗去。然后使用不含RNA酶的纯水将纯化的RNA洗脱入单独的收集管,以备长期储存,也可即用于任何实验。RNeasy技术适用于大于200 nt的RNA,因此RNA产物不包括5S rRNA、tRNAs或其他小分子量RNAs。

RNeasy 96 BioRobot 9604 Kit的规格
规格 详细信息
规格 RNeasy 96孔板配套1.2 ml Collection Microtubes
样本来源 动物或人类细胞
样本量 10–5 x 105个细胞
结合能力 每孔至多100 µg RNA
1 x 105个细胞的产量*  HeLa: 1.6 µg
LMH: 1.3 µg
COS-7: 3.1 µg
Huh7: 2.0 µg
Jurkat: 1.4 µg
洗脱体积 45–140 µl
* 不同的发育阶段和生长条件等可能导致不同的产量。

Procedure

RNeasy 96纯化产品的纯化速度和效率的关键是简洁的流程。可在96孔板细胞培养皿中培养细胞并直接裂解。将裂解物转移至RNeasy 96孔板中后,每个孔中的RNA与硅胶模结合,污染物被洗去。然后使用不含RNA酶的纯水将纯化的RNA洗脱入单独的收集管,以备长期储存,也可即用于任何实验。RNeasy 96 BioRobot 9604 Kit具有纯化总RNA和细胞质RNA的不同实验方案。可在BioRobot 9604上自动纯化RNA。如果下游应用需要,可使用RNeasy 96 Kit在QIAvac 96真空底座和/或96-Well-Plate Centrifugation System上手动纯化RNA。

使用RNeasy 96 BioRobot 8000 Kit在BioRobot 8000工作站或BioRobot Universal System上可实现完全自动纯化RNA。

Applications

RNeasy 96 BioRobot 9604 Kit和RNeasy 96 Kits纯化得到的高品质RNA可用于:

基因表达分析中的高通量RNA应用
real-time RT-PCR等灵敏的应用
药理和毒理研究
高通量药物筛选 
生物医药研究和分子诊断
Features
Specifications
应用 PCR, real-time PCR, microarray
洗脱体积 45–140 µl
自动化处理 BioRobot 9604 or BioRobot 3000 Workstation
规格 96-well plate
主要样本类型 Cultured cells
处理 Automated
纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白 RNA
样本量 5 x 10e5
技术 Silica technology
产量 1.3 –3.1 µg

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试剂盒操作手册 (1)
For automated RNA isolation from animal and human cells
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