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QIAGEN Plasmid Kits

纯化至多10 mg转染级质粒或柯斯质粒DNA

  • 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

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。

Cat No./ID: 10023
QIAGEN-tip 20 (25)
25 columns
Cat No./ID: 10043
QIAGEN-tip 100 (25)
25 columns
Cat No./ID: 10063
QIAGEN-tip 500 (25)
25 columns
Cat No./ID: 10083
QIAGEN-tip 2500 (25)
25 columns
Cat No./ID: 10091
QIAGEN-tip 10000 (5)
5 columns
Cat No./ID: 12123
QIAGEN Plasmid Mini Kit (25)
25 QIAGEN-tip 20, Reagents, Buffers
Cat No./ID: 12125
QIAGEN Plasmid Mini Kit (100)
100 QIAGEN-tip 20, Reagents, Buffers
Cat No./ID: 12143
QIAGEN Plasmid Midi Kit (25)
25 QIAGEN-tip 100, Reagents, Buffers
Cat No./ID: 12145
QIAGEN Plasmid Midi Kit (100)
100 QIAGEN-tip 100, Reagents, Buffers
Cat No./ID: 12145X4
QIAGEN Plasmid Midi Kit (400)
400 QIAGEN-tip 100, Reagents, Buffers
Cat No./ID: 12162
QIAGEN Plasmid Maxi Kit (10)
10 QIAGEN-tip 500, Reagents, Buffers
Cat No./ID: 12163
QIAGEN Plasmid Maxi Kit (25)
25 QIAGEN-tip 500, Reagents, Buffers
Cat No./ID: 12165
QIAGEN Plasmid Maxi Kit (100)
100 QIAGEN-tip 500, Reagents, Buffers
Cat No./ID: 12181
QIAGEN Plasmid Mega Kit (5)
5 QIAGEN-tip 2500, Reagents, Buffers
Cat No./ID: 12183
QIAGEN Plasmid Mega Kit (25)
25 QIAGEN-tip 2500, Reagents, Buffers
Cat No./ID: 12191
QIAGEN Plasmid Giga Kit (5)
5 QIAGEN-tip 10000, Reagents, Buffers
Cat No./ID: 19046
Plasmid Buffer Set
Buffers P1, P2, P3, QBT, QC, QF, RNase A; for 100 plasmid mini-, 25 midi-, or 10 maxipreps

QIAGEN Plasmid Kits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。


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转染效率与质粒纯化方法。
采用不同方法制备pRSVcat DNA,通过脂质体介导的转染将其导入指定的细胞系内,40小时后测定CAT表达水平,确定转染效率。每个柱形代表4个独立的转录的平均值(2个质粒制备物分别转染2次)。使用QIAGEN Plasmid Kit可获得最高的转染效率。
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QIAGEN Plasmid Kit procedures.
Neutralized bacterial lysates are cleared by centrifugation. 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. The DNA is concentrated and desalted by isopropanol precipitation and collected by centrifugation.
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(参见"Transfection efficiency versus plasmid purification method")、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(参见"QIAGEN Plasmid Kit procedures")。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
Features
Specifications
应用 Transfection, cloning, sequencing, capillary sequencing etc.
起始样本体积 2.5–5 liters culture volume
质粒类型 High-copy, low-copy, cosmid DNA
处理 Manual (centrifugation)
每次处理样本量;通量 1 sample per run
技术 Anion-exchange technology
每次运行时间或处理样本数 320 min
产量 <10 mg

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常见问题 (50)
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补充实验方案 (8)

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This protocol is designed to provide up to 150 μg BAC/PAC/P1 DNA or up to 400 μg cosmid DNA.
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This protocol is designed for the rapid, easy, and non-toxic preparation of up to 2 mg genomic DNA from not more than 2 g of tissue using QIAGEN-tip 2500. QIAGEN® Genomic-tips 20/G, 100/G, and 500/G can also be used with this protocol by reducing the amount of starting material according to the table on page 2. The purified genomic DNA ranges in size from 50-150 kb.
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Endotoxin-free DNA is essential for gene therapy research and will improve transfection into sensitive eukaryotic cells.
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This protocol is designed for isolation of up to 200 μg RNA from 150 mg plant tissue or up to 1 mg RNA from 600 mg plant tissue and is for use with QIAGEN-tip 100 or QIAGEN-tip 500, respectively.
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This protocol is for purification of up to 100 µg endotoxin-free plasmid DNA using QIAGEN-tip 100.
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用户开发的实验方案 (12)
The procedure has been used successfully for isolation of plasmids SCP2 and SCP2* as well as the plasmids listed in Table 1 (see next page) from S. coelciolor and S. lividans strains.
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This procedure has been used successfully for isolation of 150-250 kb BAC DNA from a mouse-BAC library cloned in pBeloBAC11 from Escherichia coli strain HB101/r. The yield of BAC DNA from 100 ml culture was typically 20-40 μg.

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The procedure has been used successfully for isolation of high- and low-copy-number plasmids from various Bacillus subtilis strains. Yield of plasmid DNA was typically 10-20 µg plasmid DNA from 100 ml culture.
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The procedure has been used successfully for isolation of 110 kb P1 DNA (pAdsacBII with an 80 kb insert) from Escherichia coli strain NS3529. Yield of P1 DNA was typically 10-50 µg from 500 ml culture.
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The procedure has been used successfully for isolation of linear plasmids from Borrelia burgdorferi sensu lato species, which include Borrelia burgdorferi sensu stricto, Borrelia afzelli, and Borrelia garinii.
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The procedure has been used successfully for isolation of high-copy-number plasmids from Citrobacter freundii. Yield of plasmid DNA was typically 3-8 µg DNA per ml culture.
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The procedure has been used successfully for isolation of cryptic plasmids (pLC2-based) from mesophilic Lactobacillus strains such as L. sake and L. curvatus. Yield of plasmid DNA was typically 10-20 µg plasmid DNA from 100 ml culture.
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The procedure has been used successfully for isolation of the large (128 kb), very-low-copynumber (1-2 copies per cell) plasmid pHCG3 and its derivatives from Oligotropha carboxidovorans. Yield of plasmid DNA was typically 3-6 µg plasmid DNA from 200 ml culture.
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The procedure has been used successfully for isolation of high-copy-number plasmids from Proteus vulgaris and Proteus mirabilis. Yield of plasmid DNA was typically 3-8 µg DNA per ml culture.
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The procedure has been used successfully for isolation of a variety of medium-copy-number shuttle vectors from S. xylosus, S. carnosus, S. epidermidis, and S. aureus. Yield of plasmid DNA was typically 2-10 µg from 50 ml culture.
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The procedure has been used successfully for isolation of different medium-copy-number plasmids carrying pHM1519 or pBL1 origins of replication from Corynebacterium glutamicum ATCC 13032. Yield of plasmid DNA was typically 0.4-1.5 µg per ml LB culture, although yield was dependent on the vector, the insert, and the size of the plasmid.
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参考文献
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