纯化至多10 mg转染级质粒或柯斯质粒DNA
Features
纯化得到相当于连续2次CsCl梯度离心得到的DNA纯度
高产量质粒DNA
经济的制备方法
使用LyseBlue,获得最佳的裂解效果和最大的DNA产量
Product Details
QIAGEN Plasmid Kits应用重力流阴离子交换柱纯化转染级质粒DNA。该试剂盒可纯化获得至多10 mg的质粒DNA。离心获得澄清的裂解产物和异丙醇沉淀。
Performance
QIAGEN Plasmid Kit利用重力流QIAGEN阴离子交换柱技术高效纯化质粒DNA。产量可达10 mg(Giga)、2.5 mg(Mega)、500 µg(Maxi)、或100 µg(Midi)高拷贝质粒DNA(培养体积取决于质粒拷贝数、插入片段大小、宿主菌株和培养基。)QIAGEN Plasmid Kits纯化的质粒DNA适合多种应用,如:转染、克隆和体外转录。
Principle
QIAGEN-tips内独特的阴离子交换树脂专为纯化核酸设计。它独特的分离特性,使纯化的DNA的纯度相当于连续2次CsCl梯度离心得到的DNA纯度。预装的QIAGEN-tips利用重力流原理工作,缩短了纯化质粒的手动操作时间。整套QIAGEN质粒纯化体系避免苯酚、氯仿、溴化乙锭和CsCl等有毒物质,减小对使用者和环境的危害。
Procedure
使用QIAGEN Plasmid Kits,细菌裂解物通过离心澄清。澄清后的裂解物上样到阴离子交换柱,质粒DNA在适当的低盐条件和pH条件下与树脂进行选择性结合。RNA、蛋白质、代谢产物和和其它低分子量杂质用中盐洗涤液去除,用高盐缓冲液洗脱纯质粒DNA。得到的DNA用异丙醇沉淀浓缩和脱盐后,离心收集。
Applications
QIAGEN Plasmid Kits纯化质粒DNA适合多种应用,如:
转染
克隆
PCR
体外转录
Supporting data and figures
Transfection efficiency versus plasmid purification method.
Different pRSVcat DNA preparations using the methods indicated were introduced into the indicated cell lines by liposome-mediated transfection, and the efficiencies determined by measuring CAT expression levels after 40 h. Each bar represents the mean of 4 independent transfections (2 transfections with each of 2 independent plasmid preparations). The highest transfection efficiency was achieved with QIAGEN Plasmid Kits.
Specifications
Features Specifications
Technology Anion-exchange technology
Culture volume/starting material2.5–5 liters culture volume
Yield <10 mg
Processing Manual (centrifugation)
Samples per run (throughput)1 sample per run
Time per run or prep per run 320 min
Applications Transfection, cloning, sequencing, capillary sequencing etc.
Plasmid type High-copy, low-copy, cosmid DNA