Applications
The BACMAX96™ DNA Purification Kit provides easy, reliable isolation of high-quality BAC and fosmid DNA in a 96-well format. The protocol is based on a modified alkaline-lysis procedure that incorporates proprietary enzyme technologies into an easy-to-use format. Purification starts with cell pellets obtained by centrifugation of 1.2-ml E. coli cultures grown in a 96-well plate. Lysates are cleared in the presence of BACMAX96 RNase Blend, which efficiently removes unwanted RNA. Following filtration and selective precipitation steps, Plasmid-Safe™ ATPDependent DNase is used to remove any remaining bacterial chromosomal DNA.
To maximize flexibility, culture blocks and filtration plates can be supplied by the user or purchased as part of the kit. Filtration steps can therefore be performed via vacuum or centrifugation, making the kit easy to use with standard laboratory equipment. The kit is also amenable to semi- or fully-automated purification systems.
The BACMAX96 Kit consistently yields up to 1 µg of DNA from single-copy BAC and fosmid clones and more than 2.5 µg of DNA from CopyControl™ BAC and fosmid clones induced to high-copy number. The unprecedented yields from CopyControl clones are due in part to an autoinduction protocol that was specifically designed for a high-throughput format. In this "hands-off" approach, Autoinduction Solution is added to the growth media prior to inoculation and copy number remains low until shortly before the cells are harvested, ensuring clone stability.
Benefits