For isolation of up to 20 µg, 100 µg or 500 µg high-molecular-weight DNA from blood and cultured cells

Features

Reliable isolation of high-molecular-weight DNA up to 150 kb in size

No phenol or chloroform extractions

Convenient, parallel processing of multiple samples

Product Details

Blood &; Cell Culture DNA Kits provide gravity-flow, anion-exchange tips and buffers for efficient isolation 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.

The purchase of the Genomic DNA Buffer Set (cat.no. 19060) is required for yeast and bacteria samples.

Performance

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 . 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 preparing high-quality libraries for next-generation sequencing (NGS) on different platforms and is recommended by several core facilities.

Principle

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. At the same time, other cell constituents such as proteins, carbohydrates and metabolites flow through. Purified DNA is eluted in a 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.

Procedure

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.

Applications

DNA purified with the Blood &; Cell Culture DNA Kits 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

Supporting data and figures

Genomic DNA of up to 150 kb.

Pulse-field gel electrophoresis of DNA (2 µg) purified using QIAGEN Genomic-tips. M: markers.