Further information:
QIAGEN Plasmid Purification
System
For purification of BAC, PAC, P1, or cosmid DNA, free of genomic DNA
Large-construct DNA purification with this kit follows an optimized gravity-flow procedure which yields DNA of significantly greater purity than that obtained with other commonly used methods. A unique integrated treatment with ATP-Dependent Exonuclease enables efficient removal of genomic DNA (see figure "Efficient Removal of Genomic DNA").
Following alkaline lysis of up to 500 ml culture, a unique integrated digestion step with ATP Dependent Exonuclease provided with the kit, ensures selective removal of contaminating genomic DNA (see figure "Efficient Removal of Genomic DNA") as well as nicked or damaged construct DNA. The sample is then applied to a QIAGEN-tip for DNA purification. Construct DNA binds to the resin under low-salt conditions, and impurities are washed away. Genomic DNA-free ultrapure DNA is eluted in high-salt buffer and subsequently concentrated and desalted by isopropanol precipitation.
The absence of genomic DNA contamination means that DNA quantitation is accurate and reliable, enabling sensitive experiments to be carried out under defined and reproducible conditions. The ultrapure large-construct DNA is suitable for any application, including:
Efficient Removal of Genomic DNA
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ABI PRISM 7000 (TaqMan) analysis of 1 µg samples of a 105 kb BAC DNA construct isolated using either the QIAGEN Large-Construct Kit or a standard anion exchange procedure. The lower level of contaminating genomic DNA with the QIAGEN Large-Construct Kit is indicated by the greater number of amplification cycles required for its detection. |
²Ù×÷ÊÖ²á: QIAGEN Large-Construct Kit Handbook 05/02 (PDF version, 178 KB)
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* ATP solution required for the buffer is not provided. |