QIAGEN Large-Construct Kit (10)

For purification of large-molecular-weight DNA from bacterial cultures, QIAGEN's Large-Construct Kit uses gravity-flow, anion-exchange columns to recover up to 50 μg of BAC, PAC, and P1 DNA, or up to 200 μg of cosmid DNA, free of contaminating genomic DNA. The kit is processed manually by centrifugation in a single run per prep.

A unique integrated ATP-dependent exonuclease digestion step selectively removes contaminating genomic DNA as well as nicked or damaged construct DNA, producing DNA of purity equivalent to that obtained by two successive rounds of CsCl gradient centrifugation. The procedure avoids toxic substances such as phenol, chloroform, ethidium bromide, and CsCl, and the prepacked QIAGEN-tips operate by gravity flow without running dry, minimizing hands-on time. Each prep takes 280 min and uses a culture volume of up to 500 ml.

The purified DNA is suitable for subcloning, transfection, and sequencing. The kit is supplied with QIAGEN-tip 100 or QIAGEN-tip 500 columns together with Buffers P1, P2, P3, QBT, QC, and QF, and according to the manufacturer's quick-start protocol the kit can be stored at room temperature (15–25°C).