产地：QIAGEN
货号：12243
名称：QIAfilter Plasmid Midi Kit
Principle
QIAfilter Cartridges, provided in QIAfilter, HiSpeed, and EndoFree Plasmid Kits, are special filter units designed to replace centrifugation following alkaline lysis of bacterial cells. QIAfilter Cartridges completely remove SDS precipitates and clear bacterial lysates in a fraction of the time needed for centrifugation, reducing plasmid-purification time by up to 1 hour.
QIAfilter Midi Cartridges have a syringe-format (see figure "QIAfilter Midi Cartridge") and lysates are cleared in a matter of seconds by pushing the liquid through the filter.
The unique anion-exchange resin in QIAGEN-tips is developed exclusively for the purification of nucleic acids. Its exceptional separation properties result in DNA purity equivalent or superior to that obtained by two successive rounds of CsCl gradient centrifugation. Pre-packed QIAGEN-tips operate by gravity flow and never run dry, minimizing the hands-on time required for plasmid preparation. The entire QIAGEN plasmid purification system avoids the use of toxic substances such as phenol, chloroform, ethidium bromide, and CsCl, minimizing hazard both to the user and the environment.
Applications
DNA obtained with QIAfilter Plasmid Kits provides excellent results in all applications, from cloning to transfection and automated fluorescent sequencing, including capillary sequencing.
Procedure
Neutralized bacterial lysates are incubated directly in the QIAfilter Cartridge and cleared in seconds by filtration. The cleared lysate is then loaded onto the anion-exchange tip where plasmid DNA selectively binds under appropriate low-salt and pH conditions. RNA, proteins, metabolites, and other low-molecular-weight impurities are removed by a medium-salt wash, and ultrapure plasmid DNA is eluted in high-salt buffer (see flowchart "Comparison of QIAGEN Plasmid Kit Procedures"). The DNA is concentrated and desalted by isopropanol precipitation and collected by centrifugation.
QIAfilter Plasmid Midi Kit
快速纯化最高可达100 μg的质粒或粘粒DNA，转染级别纯度
无需离心即可澄清裂解液
减少质粒纯化的操作时间
高产量的质粒DNA
使用LyseBlue，获得最佳的裂解和最大的DNA产量
QIAfilter Plasmid Midi Kit 配合 QIAfilter Midi cartridges，以过滤替代离心快速澄清细菌裂解液，并使用经验证的QIAGEN-tip 100离子交换柱高效制备质粒。
QIAfilter Plasmid Midi Kit 特性
预期产量  柱子容量 培养体积  QIAfilter 规格
多至100 µg 100 µg 25-50 ml 注射规格的Midi Cartridge
实际产量取决于质粒拷贝数、插入片段尺寸、宿主菌、培养基和培养体积。
培养体积取决于质粒拷贝数、插入片段尺寸、宿主菌和培养基。
原理
QIAfilter、HiSpeed 和EndoFree Plasmid Kits产品系列中提供的QIAfilter Cartridgesre是一种特制的过滤器，专用于替代细菌细胞碱裂解之后的离心步骤。QIAfilter Cartridges 只需离心小部分的时间就可完全去除SDS沉淀并澄清细菌裂解液，将质粒制备的时间缩短至1个小时。
QIAfilter Midi Cartridges 为注射器模式（见图 "QIAfilter Midi Cartridge"），通过推动过滤器中的液体可在数秒内澄清裂解液。
QIAGEN-tips 中独特的离子交换树脂专为核酸纯化开发，其卓越的分离特性确保DNA的纯度相当于甚至高于两次连续CsCl梯度离心的效果。预装的QIAGEN-tips 基于重力流方式，用不干枯，将质粒制备的手工操作时间降至最短。整个QIAGEN质粒纯化系统避免使用有毒物质，诸如苯酚、氯仿、ethidium bromide和氯化铯，将对用户和环境的危害降至最低。
流程
将中和的细菌裂解液直接孵育在QIAfilter Cartridge中，通过几秒的过滤澄清。然后将经澄清的裂解液装载到离子交换柱，在合时的低盐浓度和pH值条件下，质粒DNA会选择性地结合上去。通过中盐浓度的洗涤液去除RNA、蛋白、代谢物和其他小分子杂质，超纯的质粒DNA洗脱至高盐度缓冲夜中（见流程图Comparison of QIAGEN Plasmid Kit Procedures）。通过异丙醇对DNA进行浓缩并脱盐，通过离心实现收集。
产地：QIAGEN
货号：12243
名称：QIAfilter Plasmid Midi Kit
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