QIAGEN Genomic-tip 500/G
For isolation of up to 500 µg high-molecular-weight DNA from a wide range of samples
QIAGEN Genomic-tips are gravity-flow, anion-exchange tips that allow 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.
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.
QIAGEN Genomic-tips come in a range of sizes to suit every preparation need and are supplied with a comprehensive handbook containing detailed protocols for genomic DNA preparation from each sample type. Specific lysis buffers for blood, tissue, cultured cells, yeast, and bacteria, and all other necessary buffers are provided in the Genomic DNA Buffer Set which can be purchased separately.
Purified DNA has been used in direct bacterial-genome sequencing.
QIAGEN Genomic-tips 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.
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 20 minutes for bacteria, 30 minutes for blood and cultured cells, and 40 minutes for tissue and yeast.
DNA purified using the QIAGEN Genomic-tip 500/G is well suited for use in the following applications:
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