QIAGEN Plasmid Plus Kits – Plasmid DNA Extraction

快速、便捷地纯化至多1 mg转染级质粒DNA

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QIAGEN Plasmid Plus Midi Kit (25)

Cat. No. / ID: 12943

25 QIAGEN Plasmid Plus Midi Columns, Extender Tubes, Reagents, Buffers, 25 QIAfilter Midi Cartridges

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KitAccessories

QIAGEN Plasmid Plus Kit

QIAvac Holder

VacValves

Column type

Midi

Maxi

Mega

Giga

Trial Kit

Preparations

100

QIAGEN Plasmid Plus 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。

想首次尝试此解决方案吗？

立即联系我们的团队，为您的 QIAGEN Plasmid Plus Midi Kit (25) 试用版试剂盒索取报价。

获取试用试剂盒报价

特点

20分钟内快速纯化至多24个样本
利用真空装置，可同时处理多个样本
低洗脱体积确保高浓度DNA
独特的洗涤缓冲液确保较低的内毒素水平
适合多种应用

产品详情

QIAGEN Plasmid Plus Kit可超快速、大规模纯化获得至多10 mg的高纯度质粒DNA。使用真空装置可同时纯化至多24个样本，有效减少手动操作时间。低洗脱体积可获得高浓度质粒DNA，无需乙醇沉淀，即可直接应用。QIAGEN Plasmid Plus Kit含有新型的洗涤缓冲液，用于去除内毒素。纯化得到的质粒DNA适合多种应用，包括在敏感的细胞株中转染。

绩效

稳定的质量和性能

QIAGEN Plasmid Plus技术可达到与阴离子交换技术相同的性能和品质。QIAGEN Plasmid Plus纯化柱独特的结合试剂确保得到高纯度质粒DNA。QIAGEN Plasmid Plus Maxi Kit提供高产量的实验方案和额外缓冲液，可获得多达10 mg高纯度质粒DNA，无RNA、蛋白质和基因组DNA等污染物（参见" High yields are ensured with QIAGEN Plasmid Plus Kits (QPP)"和" Analytical chromatogram"）。

转染级质粒DNA，内毒素水平低

内毒素，又名脂多糖或LPS，在纯化质粒过程的裂解步骤释放，是革兰氏阴性菌如E. coli细胞膜的组成部分。内毒素包括非特异性激活免疫反应，是转染实验的非可控变量，影响实验结果和重复性，使其难以比较和解释。

该试剂提供的洗涤液将内毒素水平降至1 EU/μg DNA以下。（依宿主菌、培养基和细胞密度的不同而不同。如果细胞密度很高，内毒素水平也略高，在1–3 EU/μg DNA范围内。）这使得到的质粒DNA适合多种应用，包括在敏感细胞系中转染（参见" Endotoxin levels using QIAGEN Plasmid Plus Kits"、" Endotoxin levels using different isolation methods"和" Efficient transfection in sensitive cell lines"）。

查看图表

High yields are ensured with QIAGEN Plasmid Plus Kits (QPP).

Analytical chromatogram.

Endotoxin levels using QIAGEN Plasmid Plus Kits.

Endotoxin levels using different isolation methods.

Efficient transfection in sensitive cell lines.

原理

QIAGEN Plasmid Plus Kit进行大规模质粒纯化，方便快捷。独特设计的QIAGEN Plasmid Plus纯化柱和创新的结合试剂，使其只需简单的结合–洗涤–洗脱过程。QIAGEN Plasmid Plus Kit可在20分钟内实现快速大规模的质粒制备。无需乙醇沉淀可直接用于下游各种应用。该质粒DNA适用于要求小体积量的应用。

使用LyseBlue试剂，产量最大化

该试剂盒也包含LyseBlue指示剂，用于指示最佳的缓冲液混合状态。通过简单的颜色变化，就可避免常见的手动操作失误，防止细胞裂解不充分及SDS、基因组DNA和细胞碎片沉淀不完全。这种独特的缓冲液添加剂确保了细胞充分裂解和完全中和，使产量最大化。

程序

QIAGEN Plasmid Plus Kits提供的QIAfilter试剂条能快速澄清裂解液。一步即可将裂解液装入柱中，节约时间。使用真空装置（如QIAvac 24 Plus）抽吸澄清的裂解液及后续的洗涤缓冲液，使其流过QIAGEN Plasmid Plus离心柱。将膜洗涤和干燥后，即可通过离心获得高浓度DNA（参见""）。

查看图表

QIAGEN Plasmid Plus procedure.

应用

QIAGEN Plasmid Plus Kit纯化得到的转染级质粒DNA适合多种应用，例如：

转染多数细胞系（敏感细胞系如Huh-7）
限制性酶切
克隆
体外转录和翻译
测序

辅助数据和图表

Efficient transfection in sensitive cell lines.

Plasmid pCMVβ DNA was prepared using the indicated preparation method with standard and high-yield (HY) protocol for QIAGEN Plasmid Plus Kits. Huh-7 cells were transfected using 200 ng plasmid DNA and 0.5 μl Attractene Transfection Reagent or 300 ng plasmid DNA and 0.75 μl Attractene Transfection Reagent.

资源

Technical Information

Since QIAGEN launched the world’s first kit for purification of plasmid DNA, technologies have been developed to make plasmid DNA preparation even more convenient and reduce endotoxin levels in line with scientists’ needs in the laboratory. A lot of well-known companies have developed kits, and the choice of solutions available has never been greater. We compared popular advanced-technology kits to see their relative performance in speed, convenience of handling, yield and endotoxin levels.

Download Safety Data Sheets for QIAGEN product components.

Technical Information

FAQ

We do not recommend using a vacuum manifold for DNA extraction using the QIAamp 96 DNA Blood Kit because:

Viscosity of the lysates varies from sample to sample. Therefore, the wells of the QIAamp 96 plate are at great risk of clogging.
Use of vacuum manifold would result in incomplete removal of ethanol and salts after washing with Buffer AW2. This will affect the purity and yield of samples.

FAQ ID -9158

Please contact Nalgene for ordering. Part numbers from Nalgene:

- 10 liter waste container (Nalgene Heavy Duty Vacuum Carboy [PP], cat. no. 2226-0020)

- 20 liter waste container (Nalgene Heavy Duty Vacuum Carboy [PP], cat. no. 2226-0050)

- Quick filling/venting closure (Nalgene, cat. no. 2158-0021) [lid]

- Vacuum tubing (e.g., Nalgene 180 clear plastic vacuum tubing, cat. no.8000-0065)

FAQ ID -9156

LyseBlue reagent is provided in the following QIAGEN plasmid kits:

QIAGEN Plasmid Kits (Mini-, Midi-, Maxi-, Mega and Giga)
QIAfilter Plasmid Kits (Midi-, Maxi-, Mega and Giga)
HiSpeed Plasmid Kits (Midi- and Maxi)
EndoFree Plasmid Kit (Maxi-, Mega and Giga)
QIAprep Spin Miniprep Kit
QIAGEN Plasmid Plus Kits (Midi, Maxi, Mega and Giga)

FAQ ID -865

Endotoxin level of QIAGEN Plasmid Plus kits is in the range of 1 EU/µg DNA.

The composition of Buffer S3 is confidential. This buffer is a proprietary component of QIAGEN Plasmid Plus Kits and is an plays and important role in the alkaline lysis procedure. Buffer S3 can only be purchased in combination with QIAGEN Plasmid Plus Kits.

FAQ ID -2788

VacConnectors prevent cross-contamination and are meant to be disposed after single use.

FAQ ID -9152

Yes, the waste bottles of the QIAvac Connecting System can be autoclaved. However, please be aware that the plastic may change in color.

FAQ ID -9153

No, the two buffers cannot be interchanged. The QIAGEN Plasmid Plus chemistry is not compatible with Buffer ER from the Endofree Plasmid kits.

We do not recommend using a vacuum manifold for DNA extraction of tissues using the DNeasy Blood & Tissue Kit. Viscosity of the lysates varies from sample to sample; therefore, the DNeasy mini spin columns are at great risk of clogging.

FAQ ID -9154

We do not recommend to autoclave the QIAvac 24 Plus manifold. Doing so can result in damage to the device.

FAQ ID -9151

We do not recommend using a vacuum manifold for DNA extraction with the DNeasy 96 Blood & Tissue Kit because:

Viscosity of the lysates varies from sample to sample. Therefore, the wells of the DNeasy 96 plate are at great risk of clogging.
Use of vacuum manifold would result in incomplete removal of ethanol and salts after washing with Buffer AW2. This will affect the purity and yield of samples.

FAQ ID -9157

QIAvac Multiwell can hold plates that conform to SBS plate standards, whereas the QIAvac 96 does not. Therefore, the waste tray for QIAvac Multiwell is not the same as for our QIAvac 96.

However, the waste tray from QIAvac Multiwell will fit the QIAvac 96, but not vice versa.

FAQ ID -9159

We do not have this part available for replacement. You will need to replace your vacuum manifold. Please contact your Sales representative for a price quote.

FAQ ID -9161

FAQ ID -9160

We do not recommend using a vacuum manifold for DNA extraction of tissues using the QIAamp Blood & Tissue Kit. Viscosity of the lysates varies from sample to sample; therefore, the DNeasy mini spin columns are at great risk of clogging.

FAQ ID -9155

The composition of Buffer BB is confidential. Buffer BB is a proprietary component of QIAGEN Plasmid Plus Kits and functions as a binding buffer, adjusting the correct binding conditions after alkaline lysis, when performing the QIAGEN Plasmid Plus procedure. Buffer BB can only be purchased in combination with QIAGEN Plasmid Plus Kits and is not available separately.

FAQ ID -2790

The composition of Buffer ETR is confidential. Buffer ETR is a proprietary component of QIAGEN Plasmid Plus Kits and functions as an endotoxin removal buffer. Buffer ETR can only be purchased in combination with QIAGEN Plasmid Plus Kits.

FAQ ID -2789

The filter for QIAvac Connecting System is from Pall (part no. 4400). This is the Acro 50 filter, 0.2 µm PTFE, 1/8 inch MNPT connection. Please visit www.pall.com.

FAQ ID -9150