Blood & Cell Culture DNA Midi Kit

从血液和培养细胞中纯化100 µg高分子量DNA

  • 可靠纯化150 kb的DNA
  • 无需苯酚或氯仿抽提
  • 方便同时处理多个样本

Blood & Cell Culture DNA Kit提供重力流阴离子交换柱和缓冲液,从多种生物样本中高效纯化基因组DNA。纯化得到的DNA可长达150 kb,片段平均长度为50–100 kb。

Product Cat. no. List price:
Blood & Cell Culture DNA Midi Kit (25)
25 QIAGEN Genomic-tip 100/G, QIAGEN Protease, Buffers
13343
4.290,00 kr
Add to cart
Genomic DNA Buffer Set
Buffers, including specific lysis buffers for yeast, bacteria, cells, blood, and tissue: Y1, B1, B2, C1, G2, QBT, QC, QF; for 75 mini-, 25 midi-, or 10 maxipreps
19060
1.890,00 kr
Add to cart
The Blood & Cell Culture DNA Midi Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
  • Main Image Navi
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

QIAGEN Genomic-tip流程非常温和,导致的DNA剪切可以忽略。纯化得到的DNA片段可长达150 kb,平均长度为50–100 kb(参见"Genomic DNA of up to 150 kb")。纯化得到的DNA不含RNA 、蛋白质和代谢物等污染物,A260/A280值为1.7–1.9。

纯化得到的DNA片段长度较长,尤其适用于构建高品质DNA文库以用于不同平台的二代测序,因此受到多家核心机构的推荐。

Blood & Cell Culture DNA Midi Kit的标准产量
样本来源 产量(µg)
全血(人类)
5 ml 80–100
培养细胞(HeLa )
2 x 107 80–100

原理

Blood & Cell Culture DNA Kits中的QIAGEN Genomic-tip利用QIAGEN独有的阴离子交换技术,从多种生物样本中纯化高分子量DNA,而无需苯酚或氯仿抽提。裂解缓冲液针对不同类型样本进行优化,使得核酸酶、组蛋白、DNA结合蛋白等蛋白质以及潜在的传染性病毒等快速变性。在该缓冲液的pH值以及低盐环境中,DNA与QIAGEN交换柱上的树脂结合,蛋白质、碳水化合物、代谢物等细胞组分则随流出液流出。在高盐的洗脱液中可获得纯化的DNA。Genomic-tip基于重力流技术,无需专人照看。该操作最大程度上节约了手动操作时间,是对多样本进行同时处理的理想选择。

操作流程

首先裂解样本(组织样本需要机械破碎),蛋白质在适宜的裂解缓冲液中变性(参见"QIAGEN Genomic-tip procedure")。然后加入QIAGEN Protease或蛋白酶K,温育后,将裂解产物上样到QIAGEN Genomic-tip。DNA吸附到交换柱上,其他细胞组分随流出液流出。随后进行洗涤,将残留的污染物洗去。最后洗脱得到纯化的高分子量DNA,在异丙醇中沉淀。针对血液和培养细胞的完整流程中手动操作仅需30分钟。

Blood & Cell Culture DNA Kits是即用型试剂盒,含有从全血和培养细胞中纯化高分子量DNA所需的全部组分。

应用

Blood & Cell Culture DNA Midi Kit纯化得到的DNA非常适用于以下应用:

  • Sanger测序和二代测序
  • RFLP分析
  • 靶基因分析
  • 转基因动物的筛查
  • DNA图谱研究
  • PCR扩增
Feature
Specifications
Applications PCR, RFLP, blotting, screening
Elution volume 0.1–2 ml
Format Genomic tip
Main sample type Blood, cells
Processing Manual
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein DNA
Sample amount 5 ml/2 x 10e7
Technology Anion-exchange technology
Yield 80–100 µg

You are not authorized to download the resource

Safety Data Sheets
2
Download Safety Data Sheets for QIAGEN product components.
View
Download Safety Data Sheets for QIAGEN product components.
View
Images
QIAGEN Genomic-tip procedure
QIAGEN Genomic-tip procedure.
Samples are first lysed (tissue samples are mechanically disrupted) and proteins simultaneously denatured in the appropriate lysis buffer. 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.
Genomic DNA of up to 150 kb
Genomic DNA of up to 150 kb.
Pulse-field gel electrophoresis of DNA (2 µg) purified using QIAGEN Genomic-tips. M: markers.