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Features
- Minimal mechanical shearing allows 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
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
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 (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 (see figure " Direct sequencing of bacterial genomic DNA").
QIAGEN Genomic-tip specifications
| Sample source | Amount | Yield (µg) | ||||
| Genomic-tip 20/G | Genomic-tip 100/G | Genomic-tip 500/G | Genomic-tip 20/G | Genomic-tip 100/G | Genomic-tip 500/G | |
| Whole blood (human) | 1 ml | 5 ml | 20 ml | 15–20 | 80–100 | 350–400 |
| Cultured cells (HeLa) | 5 x 106 | 2 x 107 | 1 x 108 | 15–20 | 80–100 | 350–450 |
| Tissue (liver) | 15 mg | 80 mg | 350 mg | 15–20 | 80–100 | 350–450 |
| Yeast (S. cerevisiae) | 1 ml | 5 ml | 20 ml | 18–20 | 85–95 | 350–450 |
| Gram– bacteria (E. coli) | 0.6–1.2 ml | 3–6 ml | 15–30 ml | 16–20 | 85–95 | 300–400 |
| Gram+ bacteria (B. subtilis) | 1.2–1.8 ml | 6–9 ml | 30–45 ml | 16–20 | 85–95 | 300–400 |
See figures
Principle
QIAGEN Genomic-tips use unique QIAGEN anion-exchange technology for high-molecular-weight DNA isolation 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 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 20 minutes for bacteria, 30 minutes for blood and cultured cells and 40 minutes for tissue and yeast.
See figures
Applications
DNA purified with QIAGEN Genomic-tips is well suited for use in the following applications:
- RFLP analysis
- Analysis of gene targeting
- Screening of transgenic animals
- DNA fingerprinting studies
- Southern blotting
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.
Resources
User-Developed Protocols (7)
Safety Data Sheets (4)
クイックスタートプロトコール (2)
Supplementary Protocols (1)
機器テクニカル資料 (1)
キットハンドブック (1)
MSDS (1)
Kit Handbooks (1)
Quick-Start Protocols (2)
Instrument Technical Documents (1)
Publications
Similarity and differences in the Lactobacillus acidophilus group identified by polyphasic analysis and comparative genomics.
J Bacteriol; 2006; 189 (4):1311-21 2006 Dec 1 PMID:17142402
A genome-wide map of diversity in Plasmodium falciparum.
Nat Genet; 2006; 39 (1):113-9 2006 Dec 10 PMID:17159979
Development and evaluation of a loop-mediated isothermal amplification method for rapid diagnosis of Bordetella pertussis infection.
J Clin Microbiol; 2006; 44 (5):1899-902 2006 May PMID:16672435
Arsenic detoxification and evolution of trimethylarsine gas by a microbial arsenite S-adenosylmethionine methyltransferase.
Proc Natl Acad Sci U S A; 2006; 103 (7):2075-80 2006 Feb 1 PMID:16452170
Validation of a 21-locus autosomal SNP multiplex for forensic identification purposes.
Forensic Sci Int; 2005; 154 (1):62-77 2005 Jan 20 PMID:16182950
FAQ
Do you have a protocol for isolation of genomic DNA from plants and filamentous fungi using QIAGEN Genomic-tips?
Do you have a protocol for isolation of genomic DNA from flies using QIAGEN Genomic-tips?
Do you have a protocol for isolation of genomic DNA from insects?
Do you have a protocol for the purification of DNA from fruit flies?
Do you have a protocol for isolation of genomic and plasmid DNA from cell cultures using QIAGEN Genomic-tips?
Do you have a protocol for the isolation of genomic DNA from frozen clotted blood?
Do you have a protocol for the isolation of genomic DNA from fungi?
Do you have protocols for the isolation of genomic DNA from tissue using QIAGEN-tips 2500 (Mega) and 10000 (Giga)?
Do you have a protocol for isolation of genomic and viral DNA from lymphocytes using QIAGEN Genomic-tips?
What do you recommend for the cleanup of genomic DNA (gDNA)?
What is the size of genomic DNA that is obtained with QIAGEN Genomic-tips?