QIAGEN Large-Construct Kit
纯化至多50 μg BAC、PAC和P1 DNA或至多200 μg的柯斯质粒DNA和非基因组DNA
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QIAGEN Large-Construct Kit (10)
目录编号 / ID. 12462
10 QIAGEN-tip 500, Reagents, Buffers, ATP-Dependent Exonuclease
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QIAGEN Large-Construct Kit 旨在用于分子生物学应用。该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
- 高效去除基因组DNA污染
- 转染级别纯度的高分子量DNA
- 快速简单的操作
产品详情
QIAGEN Large-Construct Kit提供重力流和阴离子交换柱，用于纯化大分子量的DNA。独特的ATP-dependent外切酶消化步骤，确保特异性去除基因组DNA。产物DNA的纯度相当于经过两次CsCl密度梯度离心得到的纯度，适用于转染级别的应用。
绩效
使用QIAGEN Large-Construct Kit纯化DNA，无基因组DNA的污染（参见" Efficient removal of genomic DNA"）。基因组DNA的去除确保准确可靠的进行DNA定量，使一些敏感实验可在明确的和可重复的条件下进行。
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原理
QIAGEN Large-Construct Kit使用经优化的重力流操作，获得的DNA的纯度显著高于其他常规方法获得的DNA的纯度。独特的ATP-dependent外切酶消化步骤，确保特异性去除基因组DNA。
QIAGEN Large-Construct Kit含有的QIAGEN-tips中独特的阴离子交换树脂专为核酸纯化设计。其出色的核酸分离特性可纯化得到高纯度DNA，相当于经过两次氯化铯梯度离心得到的纯度。预装的QIAGEN-tips（参见" Anion-exchange tips"）运用重力沉降原理，减少了质粒制备过程所需的手工操作时间。整个QIAGEN质粒纯化系统不使用苯酚、氯仿、溴乙锭和氯化铯等有毒物质，最大程度减小了对使用者及环境的危害。
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程序
使用碱裂解法裂解多达500 ml的培养液后（参见" QIAGEN Plasmid Kit procedures"），使用试剂盒中的ATP-dependent外切酶进行消化，确保特异性去除基因组DNA及缺损的高分子量DNA。之后将样本上样至阴离子交换柱，在适当的低盐浓度和pH值条件下，质粒DNA与树脂特异性结合。用中等盐浓度的缓冲液洗涤，去除RNA、蛋白质、代谢物和其他小分子的杂质。用高盐缓冲液洗脱已去除基因组DNA的纯化质粒DNA。加入异丙醇使DNA浓缩和脱盐，之后离心收集。
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应用
QIAGEN Large-Construct Kit纯化获得的DNA适用于各种下游应用，包括：
- 亚克隆和鸟枪法制备DNA文库
- 对高分子量DNA直接进行测序
- 转染
辅助数据和图表
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.
规格
| 特点 | 规格 |
|---|---|
| plasmidtype | BAC, PAC, P1, cosmid DNA |
| applications | Subcloning, transfection, sequencing etc. |
| processing | Manual (centrifugation) |
| culturevolumestartingmaterial | <500 ml culture volume |
| samplesperrunthroughput | 1 sample per run |
| technology | Anion-exchange technology |
| timeperrunorprepperrun | 280 min |
| yield | <150 ug |
资源
快速启动实验方案 (2)
试剂盒操作手册 (1)
安全数据表 (1)
Certificates of Analysis (1)
Kit Handbooks (1)
Safety Data Sheets (1)
Quick-Start Protocols (2)
文献
Targeted disruption of Kaposi's sarcoma-associated herpesvirus ORF57 in the viral genome is detrimental for the expression of ORF59, K8alpha, and K8.1 and the production of infectious virus.
J Virol; 2006; 81 (3):1062-71 2006 Nov 15 PMID:17108026
Functional characterization of Kaposi's sarcoma-associated herpesvirus ORF45 by bacterial artificial chromosome-based mutagenesis.
J Virol; 2006; 80 (24):12187-96 2006 Oct 11 PMID:17035322
Autographa californica multiple nucleopolyhedrovirus nucleocapsid assembly is interrupted upon deletion of the 38K gene.
J Virol; 2006; 80 (23):11475-85 2006 Sep 20 PMID:16987976
Identification of in vivo-induced bacterial protein antigens during human infection with Salmonella enterica serovar Typhi.
Infect Immun; 2006; 74 (9):5161-8 2006 Sep PMID:16926408
Molecular characterization of a diagnostic DNA marker for domesticated tetraploid wheat provides evidence for gene flow from wild tetraploid wheat to hexaploid wheat.
Mol Biol Evol; 2006; 23 (7):1386-96 2006 May 4 PMID:16675504