QIAquick 96 PCR Purification Kit

96孔板纯化96个PCR产物(多至10 μg),100 bp–10 kb

S_1346_DNA_QQ0827

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QIAquick 96 PCR Purification Kit (4)

Cat. No. / ID:  28181

用于 4 次 96 PCR 反应的纯化:4 个 QIAquick 96 Plates、缓冲液、采样微量管 (1.2 ml)、盖子
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Kit
QIAquick 96 PCR Purification Kit
QIAquick 96 PCR BioRobot Kit
Preparations
4
24
QIAquick 96 PCR Purification Kit 旨在用于分子生物学应用。这些产品不能用于疾病诊断、预防和治疗。

✓ 全天候自动处理在线订单

✓ 博学专业的产品和技术支持

✓ 快速可靠的(再)订购

特点

  • 即用型DNA回收率高达95%
  • 流程快速、方便
  • 3个简单步骤即可回收多达10 kb DNA片段

产品详情

QIAquick 96 PCR Purification Kits含有96孔板、缓冲液和收集管,利用硅胶模纯化技术高通量纯化大于100 bp PCR产物。通过简单、快速的结合、洗涤、洗脱的流程纯化长达10 kb的DNA,洗脱体积为60–80 μl(最终洗脱体积为40–60 μl)。回收过程可在BioRobot工作站上使用QIAquick 96 PCR BioRobot Kit全自动运行。

绩效

QIAquick 96 PCR Kit提供快速、简单的方法,高通量纯化样本,回收率高达90%。QIAquick 96操作流程提供高纯度DNA适用于不同的下游应用(参见"Accurate sequencing")。采用QIAvac 96,可从1–4 x 96个样本中纯化得到100 bp–10 kb的DNA片段。

原理

QIAquick 96 Kits包含一个硅胶膜,可在高盐缓冲液中结合DNA,并且在低盐缓冲液或水中洗脱。纯化过程去除引物、核苷酸、酶、矿物油、盐、琼脂糖、溴化乙锭和DNA样本中的其他杂质。硅胶膜技术结局了与树脂松动的问题和不便。优化特殊的结合缓冲液用于特殊应用,并且在特定的片段范围内,促进DNA的选择性吸附。

QIAquick 96操作流程平行纯化多达96个样本,并在QIAvac 96上采用高效真空驱动的纯化过程。

程序

QIAquick体系采用简单的结合、洗涤、洗脱操作流程(参见"QIAquick 96 procedure")。将结合缓冲液直接加到PCR样本或者其他酶试剂中,然后将混合液上样到96孔板上。在高盐的缓冲液中,核酸吸附到硅胶膜上。洗去杂质,并且用少量的低盐缓冲液或纯水洗脱纯DNA,为后续应用准备。

处理过程

QIAquick多孔模块在QIAvac底座上采用真空驱动的纯化过程处理。QIAquick 96 PCR Purification Kit需要使用QIAvac 96真空底座。产物回收过程可以使用QIAquick 96 PCR BioRobot Kit在BioRobot工作站上自动运行。

应用

采用MinElute或者QIAquick体系纯化的DNA片段可即用于多种下游应用,包括测序、微阵列分析、连接和转化、限制性酶切、标记、显微注射、PCR和体外转录。

辅助数据和图表

Specifications

FeaturesSpecifications
Binding capacity10 µg
Processing手动/自动
Removal <10mers 17–40mers dye terminator proteins去除 <40mer
Sample type: applicationsDNA、寡核苷酸:PCR 反应
Format96 孔板
Fragment size100 bp – 10 kb
Technology硅胶膜技术
Recovery: oligonucleotides dsDNA回收率:寡核苷酸、dsDNA
Elution volume60-80 µl

资源

快速启动实验方案 (1)
Technical Information and Important Notes (2)
试剂盒操作手册 (1)
For rapid purification of multiple PCR products 
安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
Safety Data Sheets (1)
Certificates of Analysis (1)

Publications

Temporal and differential gene expression of Singapore grouper iridovirus.
Chen LM; Wang F; Song W; Hew CL;
J Gen Virol; 2006; 87 (Pt 10):2907-2915 2006 Oct PMID:16963749
Comparative genomics of host-specific virulence in Pseudomonas syringae.
Sarkar SF; Gordon JS; Martin GB; Guttman DS;
Genetics; 2006; 174 (2):1041-56 2006 Sep 1 PMID:16951068
cDNA microarrays as a tool for identification of biomineralization proteins in the coccolithophorid Emiliania huxleyi (Haptophyta).
Quinn P; Bowers RM; Zhang X; Wahlund TM; Fanelli MA; Olszova D; Read BA;
Appl Environ Microbiol; 2006; 72 (8):5512-26 2006 Aug PMID:16885305
Characterization of the vernalization response in Lolium perenne by a cDNA microarray approach.
Ciannamea S; Busscher-Lange J; de Folter S; Angenent GC; Immink RG;
Plant Cell Physiol; 2006; 47 (4):481-92 2006 Jan 31 PMID:16449231
Anti-inflammatory activity in vitro and in vivo of the protein farnesyltransferase inhibitor tipifarnib.
Xue X; Lai KT; Huang JF; Gu Y; Karlsson L; Fourie A;
J Pharmacol Exp Ther; 2005; 317 (1):53-60 2005 Dec 13 PMID:16352705

FAQ

I bound an 11 kb DNA fragment to a QIAquick column; is it completely lost?

Larger DNA fragments bind more tightly to the QIAquick columns. It is difficult to predict whether a DNA fragment larger than 10 kb can be efficiently recovered, because this depends on base composition as well as fragment size. If the fragment is only a few kb larger than the 10 kb limit, it can be helpful to heat the elution buffer EB to 60°C and let it incubate on the column for a few minutes before centrifuging. However, please note that it will become less likely to recover your sample the larger the fragment size is. As we cannot guarantee recovery of fragments larger than the maximum cutoff size, we do not recommend to purify such fragments using QIAquick Kits.

The QIAEX II Kit can be used to extract DNA fragments up to 50 kb from agarose or polyacrylamide gels.

 

FAQ ID -756
How can I improve recoveries when using the QIAquick Kits?

Buffer PE did not contain ethanol

Ethanol must be added to Buffer PE (concentrate) before use. Repeat procedure with correctly prepared Buffer PE.

Inappropriate elution buffer

DNA will only be eluted efficiently in the presence of low-salt buffer (e.g., Buffer EB: 10 mM Tris·Cl, pH 8.5) or water. Elution efficiency is strongly dependent on the salt concentration and pH of the elution buffer. Contrary to adsorption, elution is most efficient under basic conditions and low salt concentrations. DNA is eluted with 50 or 30 µl of the provided Buffer EB (10 mM Tris·Cl, pH 8.5), or water. The maximum elution efficiency is achieved between pH 7.0 and 8.5. When using water to elute, make sure that the pH is within this range. In addition, DNA must be stored at –20°C when eluted with water since DNA may degrade in the absence of a buffering agent. Elution with TE (10 mM Tris·Cl, 1 mM EDTA, pH 8.0) is possible, but not recommended because EDTA may inhibit subsequent enzymatic reactions.

Elution buffer incorrectly dispensed

Add elution buffer to the center of the QIAquick membrane to ensure that the buffer completely covers the membrane. This is particularly important when using small elution volumes (30 µl).

FAQ ID -180
Can I use a centrifuge instead of vacuum when using the QIAquick 96 PCR Purification Kit?
Yes, you can use a QIAGEN Centrifuge with the QIAquick 96 PCR Purification Kit. Follow the Supplementary Protocol 'Spin procedure for purifying 2 x 96 PCR samples using the Plate Rotor 2 x 96, a special centrifuge, and the QIAquick 96 PCR Purification Kit.' Here's the link to the protocol.
FAQ ID -293
Do you have a protocol for purifying 2x96 PCR samples simultaneously with the QIAquick 96 PCR Purification Kit?

Yes, please follow the Supplementary Protocol 'Spin procedure for purifying the 2x96 PCR samples using the Plate Rotor 2x96, a special centrifuge and the QIAquick 96 PCR Purification Kit' (QQ01).  Please contact your local QIAGEN Technical Service for this protocol.

The protocol is for use with QIAGEN Centrifuges and the Plate Rotor 2 x 96.

FAQ ID -935
Do I have to remove the oil from my PCR reaction before using the QIAquick or MinElute PCR Purification Kit?
No - mineral oil will not affect the clean-up procedure with the QIAquick or MinElute PCR Purification Kit.
FAQ ID -575
Do you have protocols for multiple extractions of DNA fragments from agarose gels?

Yes, please follow the Supplementary Protocols 'High-throughput gel extractions using the QIAquick 96 PCR Purification Kit' (QQ03).  Please contact your local QIAGEN Technical Service for this protocol.

FAQ ID -944
Why does my DNA sample float out of the slot when loading it onto an agarose gel?

DNA fragments purified with the QIAGEN DNA Cleanup Systems, i.e., the QIAquick PCR Purification Kit, the MinElute Reaction Cleanup Kit, the QIAEX II Gel Extraction Kit etc. may float out of the loading wells of agarose gels due to residual ethanol carried over from the wash step with Buffer PE (despite the addtition of glycerol-containing loading buffer).

Use either of the following options to remove residual ethanol from the eluate:

  • re-purify the sample using a QIAquick-, or MinElute column, or QIAEX II resin
  • incubate the eluate at 56°C for 10 min to evaporate the ethanol
  • dry down the sample in a vacuum centrifuge, and resuspend the pellet in a small volume of sterile water
FAQ ID -205