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RNeasy 96 QIAcube HT Kit

自动、高通量从人类和动物细胞、组织中分离RNA
  • 简便、可靠的自动处理,节省时间和费用
  • 灵活的流程可以分离RNA、或包括miRNA在内的总RNA
  • 适用于多种起始材料,提供针对不同大小RNA的实验方案
  • 高品质的纯RNA可即用于多种下游应用
  • 从少量起始样本中可获得稳定产量的RNA
RNeasy 96 QIAcube HT Kit能够在QIAcube HT仪器上,从动物或人类细胞和组织样本中,便利的自动纯化RNA,或包含miRNA在内的总RNA。该试剂盒采用成熟的RNeasy硅胶模技术和便利的96孔板规格,能够去除抑制剂和污染物,纯化的高品质RNA即用于下游分析。
Cat No./ID: 74171
RNeasy 96 QIAcube HT Kit (5)
For 480 preps: RNeasy 96 plates, RNase-free water, buffers
Cat No./ID: 950067
QIAcube HT Plasticware
Plasticware for 480 typical preps on QIAcube HT:
5 S-Blocks, 5 Elution Microtubes RS (EMTR), 120 x 8-Well Strip Caps for EMTR, 9 x 96 Filter-Tips OnCor C, TapePad

RNeasy 96 QIAcube HT Kit适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


0
从组织中自动纯化RNA。
从多个10 mg的(RNAlater RNA Stabilization Reagent稳定的)大鼠组织中分离RNA。使用QIAzol Lysis Reagent和QIAcube HT、BioRobot Universal System或QIAcube全自动核酸纯化仪纯化RNA。之后使用QuantiFast Probe RT-PCR Kit通过qRT-PCR分析pgk1 mRNA。CT值显示不同纯化平台纯化的RNA具有相似表现。
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从组织中纯化高品质RNA。
从多个10 mg的(RNAlater RNA Stabilization Reagent稳定的)大鼠肺、肾、心、大脑、脾和肠组织中分离RNA。使用QIAzol Lysis Reagent和QIAcube HT、BioRobot Universal System或QIAcube全自动核酸纯化仪纯化RNA。之后从每份纯化RNA中取1 µl,使用QIAxcel RNA QC Kit v2.0和QX RNA Size Marker 200–6000 nt在QIAxcel Advanced上进行分析。RIS值显示纯化RNA都具有高品质。
Performance
在QIAcube HT上使用RNeasy 96 QIAcube HT Kit纯化RNA十分便利、可靠,在96孔板上无明显的孔间差异(参见Linear RNA recovery and Reproducible 96-well RNA purification)。

这一经济、易用的产品纯化的RNA品质与其他备受信赖的QIAGEN纯化试剂盒相同(参见Automated purification of RNA from tissues)。使用QIAxcel Advanced对纯化产物RNA进行分析,其RIS(RNA完整度值)高,在实验中的表现与其他QIAGEN纯化方案所获得的产物相当(参见High-quality RNA from tissues)。

RNeasy 96 QIAcube HT Kit从少量起始材料中纯化的RNA产量稳定(参见下表)。

RNeasy 96 QIAcube HT Kit纯化的总RNA的常规产量
细胞系 来源 RNA产量
(µg /105 个细胞)*
HeLa 人子宫颈癌细胞 1.6
LMH 鸡肝癌细胞 1.3
COS-7 猴肾,SV-40 transformed 3.1
Huh7 人肝癌细胞 2.0
Jurkat 人T细胞白血病 1.4
K-562 人慢性粒细胞白血病 1.9
* 不同物种、发展阶段和生长条件会导致不同产量。如果RNeasy实验流程富集的是 >200个核苷酸的RNA,总RNA产物中不包含5S rRNA、tRNA或其他低分子量RNA(占细胞总RNA的15–20%)。
Principle
RNeasy 96 QIAcube HT Kit能够在QIAcube HT仪器上,从5 x 105个动物或人类细胞或40 mg以内的组织中自动纯化RNA或包含miRNA在内的总RNA。依据不同起始材料和所需产物大小,提供6种实验方案。细胞样本可以直接使用RNeasy 96 QIAcube HT Kit和专用的实验方案进行纯化。 组织样本(<40 mg)和需要纯化小RNA的样本需要先在QIAzol Lysis Reagent中裂解。纯化产物为高品质RNA,在众多下游应用中表现出色。

RNeasy 96 QIAcube HT Kit结合了硅胶模的选择性结合特性和96孔板的高通量优点,可在QIAcube HT仪器上全自动同时处理24–96个样本。 

从细胞中纯化总RNA的规格
规格 描述
样本数量 24–96个样本(每批8个)
样本起始体积 最多5 x 105个细胞,需在140 μl Buffer RLT中匀浆
洗脱体积 110 μl
时间长度 大约93或114*分钟内处理96个样本
大约54或73*分钟内处理24个样本
* 包括可选的DNase消化步骤。

从组织中纯化总RNA的规格
规格 描述
样本数量 24–96个样本(每批8个)
样本起始体积 从40 mg冰冻的活20 mg稳定的组织(在750 μl QIAzol Lysis Reagent中匀浆)中获得的350 μl水相
洗脱体积 110 μl
时间长度 大约73或93*分钟内处理96个样本
大约34或54*分钟内处理24个样本
* 包括可选的DNase消化步骤。

从细胞或组织中纯化包含miRNA在内的总RNA
规格 描述
样本数量 24–96个样本(每批8个)
样本起始体积 从40 mg冰冻的活20 mg稳定的组织(在750 μl QIAzol Lysis Reagent中匀浆)中获得的350 μl水相
洗脱体积 110 μl
时间长度 大约78分钟内处理96个样本
大约38分钟内处理24个样本

Procedure
在高度易变性条件下,用异硫氰酸胍使RNase失活,手动裂解细胞,确保纯化得到完整的RNA。开始操作后,在裂解液中加入乙醇,达到合适的结合条件,然后将样本加入RNeasy 96孔板。总RNA和污染物被高效洗去。之后用少量的不含RNase的水洗脱获得高品质RNA,可即用于多种下游应用。

按照从动物或人类细胞中纯化总RNA的自动实验方案进行操作,可纯化获得长于200个核苷酸的RNA。此方法可富集mRNA,因为多数RNA小于200个核苷酸(例如5.8S rRNA、5S rRNA和tRNA,其总共占总RNA的15–20%)已被选择性去除。纯化的RNA的长度与CsCl离心纯化法(笑RNA不会沉淀下来)得到的RNA长度相当。

如需从组织中纯化总RNA,或从细胞和组织中纯化包含小RNA在内的总RNA,我们推荐使用QIAzol Lysis Reagent裂解。详细实验方案请参见试剂盒说明书。
Applications
RNeasy 96 QIAcube HT Kit可高通量、高效的纯化RNA,可用于药物研究和科研等多领域。纯化的RNA即用于多种下游应用,包括:
  • RT-PCR和qRT-PCR
  • 差异显示 
  • cDNA合成
  • Northern印迹、点印迹和狭缝印迹
  • Pri引物延伸
  • Poly A+ RNA选择
  • RNase/S1核糖核酸酶保护

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产品介绍与指南 (1)
For fast and reliable automated 96-well nucleic acid purification
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实验方案软件 (14)
Q Protocol notes for purification of total RNA or total RNA, including miRNA, from animal tissue and cells on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the miRNeasy 96 QIAcube HT V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Installation file (.msi) for installing the Q Protocol run file (.qsp) in the QIAcube HT software. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Q Protocol notes for automatic transfer of upper aqueous phase to an empty S-Block on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT pre-treatment V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Installation file (.msi) for installing the Q Protocol run file (.qsp) in the QIAcube HT software, For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Q Protocol notes for purification of total RNA from animal or human cells, including DNase digestion, on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT total RNA Cell with DNase V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Q Protocol notes for purification of total RNA from animal or human cells on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT total RNA Cell V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Installation file (.msi) for installing the Q Protocol run file (.qsp) in the QIAcube HT software. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Installation file (.msi) for installing the Q Protocol run file (.qsp) in the QIAcube HT software. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Q Protocol notes for purification of total RNA from tissue, including DNase digestion, on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT total RNA Tissue with DNase V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Q Protocol notes for purification of total RNA from tissue on the QIAcube HT using the RNeasy 96 QIAcube HT Kit; use with the RNeasy 96 QIAcube HT total RNA Tissue V1.qsp run file. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Installation file (.msi) for installing the Q Protocol run file (.qsp) in the QIAcube HT software. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Installation file (.msi) for installing the Q Protocol run file (.qsp) in the QIAcube HT software. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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Procedure for installing Q Protocols for QIAcube HT Kits on the QIAcube HT and the QIAxtractor. For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software.
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These Q Protocols and installation files can only be used with QIAcube HT Operating software version 4.17 and are not compatible with the new QIAcube HT Prep Manager software.

QIAcube HT Prep Manager Software already includes all standard protocols. Additional protocols and kit configurations can be easily installed. Check for available updates under the "Operating Software" section of the "Product Resources" tab.
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用户使用手册 (2)
Instrument user manual for automated mid- to high-throughput nucleic acid purification in 96-well format using silica membrane technology; for use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software
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Instrument user manual for automated mid- to high-throughput nucleic acid purification in 96-well format using silica membrane technology; for use with QIAcube HT Prep Manager Software.
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快速启动实验方案 (4)
For use with the RNeasy 96 QIAcube HT Kit and QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software
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For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software
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For use with QIAcube HT Operating Software 4.17; not for use with QIAcube HT Prep Manager Software
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For use with QIAcube HT Prep Manager Software; not compatible with QIAcube HT Operating Software 4.17
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试剂盒操作手册 (2)
For use with QIAcube HT Prep Manager Software; not compatible with QIAcube HT Operating Software 4.17
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Handbook for purification of total RNA, or total RNA, including miRNA, from animal tissue and cells on the QIAcube HT using the RNeasy 96 QIAcube HT Kit. For use with QIAcube HT Operating Software 4.17; not compatible with QIAcube HT Prep Manager Software.
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