Blood & Cell Culture DNA Mini Kit

For isolation of up to 20 µg high-molecular-weight DNA from blood and cultured cells
  • Reliable isolation of DNA up to 150 kb in size
  • No phenol or chloroform extractions
  • Convenient, parallel processing of multiple samples

Blood & Cell Culture DNA Kits provide gravity-flow, anion-exchange tips and buffers for the efficient purification of genomic DNA from a wide range of biological samples. The purified DNA is sized up to 150 kb with an average size of 50–100 kb.

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产品 货号 目录价:
Blood & Cell Culture DNA Mini Kit (25)
25 QIAGEN Genomic-tip 20/G, QIAGEN Protease, Buffers
13323
2.260,00 kr
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Blood & Cell Culture DNA Mini Kit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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QIAGEN Genomic-tip实验流程。|长达150 kb的基因组DNA。|
先裂解样本(组织样本采用机械破碎),同时将蛋白质置于适当的裂解缓冲液中变性。然后加入QIAGEN Protease或Proteinase K,孵育一段时间后,将裂解物上样至QIAGEN Genomic-tip。DNA结合至离心柱上,其他细胞成分则可通过。通过洗涤步骤去除残留的污染物后,洗脱纯的高分子量DNA,并利用异丙醇进行沉淀。|通过脉冲场凝胶电泳对QIAGEN Genomic-tips纯化获得的DNA(2 µg)进行电泳分析。M: 标记物。|
性能

The QIAGEN Genomic-tip procedure is very gentle and results in negligible DNA shearing. DNA purified with QIAGEN Genomic-tips is sized up to 150 kb with an average length of 50–100 kb (see figure "Genomic DNA of up to 150 kb"). The DNA is free of all contaminants such as RNA, protein, and metabolites, and has A260/A280 ratios between 1.7 and 1.9.

The exceptionally large size of the obtained DNA makes it especially suitable for the preparation of high-quality libraries for next generation sequencing (NGS) on different platforms and is recommended by several core facilities.

Typical yields using the Blood & Cell Culture DNA Mini Kit
Sample source Yield (µg)
Whole blood (human)

1 ml

15–20
Cultured cells (HeLa)
5 x 106 15–20

原理

QIAGEN Genomic-tips, included in Blood & Cell Culture DNA Kits, use unique QIAGEN anion-exchange technology to purify high-molecular-weight DNA from a wide range of biological samples without phenol or chloroform. Lysis buffers are optimized for different sample types and provide immediate denaturation of proteins such as nucleases, histones, and DNA-binding proteins, as well as potentially infectious viral particles. Under the pH and low-salt conditions provided by the buffer, DNA binds to the QIAGEN Resin in the column while other cell constituents such as proteins, carbohydrates, and metabolites flow through. Purified DNA is eluted in high-salt buffer. Genomic-tips operate by gravity flow, and can be left unattended without running dry. This reduces hands-on time to a minimum and makes the procedure ideal for simultaneous processing of multiple samples.

操作流程

Samples are first lysed (tissue samples are mechanically disrupted) and proteins simultaneously denatured in the appropriate lysis buffer (see flowchart "QIAGEN Genomic-tip procedure"). QIAGEN Protease or Proteinase K is then added and after a suitable incubation period, lysates are loaded onto the QIAGEN Genomic-tip. DNA binds to the column while other cell constituents pass through. Following a wash step to remove any remaining contaminants, pure, high-molecular-weight DNA is eluted and precipitated with isopropanol. Hands-on time for the complete procedure is just 30 minutes for blood and cultured cells.

Blood & Cell Culture DNA Kits are ready-to-use kits containing all the necessary components for purification of high-molecular-weight DNA from blood and cultured cells.

应用

DNA purified with the Blood & Cell Culture DNA Mini Kit is well suited for use in the following applications:

Sanger and next generation sequencing
RFLP analysis
Analysis of gene targeting
Screening of transgenic animals
DNA fingerprinting studies
PCR amplification
特点
参数
应用 PCR, RFLP, blotting, screening
洗脱体积 0.1–2 ml
规格 Genomic tip
主要样本类型 Blood, cells
处理 Manual
纯化总RNA、miRNA、poly A+ mRNA、DNA或蛋白 DNA
样本量 1 ml/5 x 10e6
技术 Anion-exchange technology
产量 15–20 µg

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参考文献
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图片
QIAGEN Genomic-tip procedure
QIAGEN Genomic-tip实验流程。
先裂解样本(组织样本采用机械破碎),同时将蛋白质置于适当的裂解缓冲液中变性。然后加入QIAGEN Protease或Proteinase K,孵育一段时间后,将裂解物上样至QIAGEN Genomic-tip。DNA结合至离心柱上,其他细胞成分则可通过。通过洗涤步骤去除残留的污染物后,洗脱纯的高分子量DNA,并利用异丙醇进行沉淀。
Genomic DNA of up to 150 kb
长达150 kb的基因组DNA。
通过脉冲场凝胶电泳对QIAGEN Genomic-tips纯化获得的DNA(2 µg)进行电泳分析。M: 标记物。