|Genome Mapping Section
DOE Human Genome Program Contractor-Grantee Workshop
58. Automated Purification of Blood, or Bacterial Genomic DNA
Dan P. Langhoff, Tuyen Nguyen, and William
In Phase I of the SBIR research we have developed a prototype of a fully automatic high-throughput blood or bacteria genomic DNA isolation instrument. Unlike any other process currently used for genomic DNA isolation, this instrument uses a derivative of electrophoretic separation technology that was developed by our company for automated purification of plasmid DNA. The separation technique is novel and powerful in that it requires no moving parts and can be performed with the combination of a simple disposable sample cassette and an inexpensive processing instrument. The process purifies high molecular weight genomic DNA direclty from a cell lysate through the use of electrophoretic movement of the DNA which is placed in between barriers of agarose medium that are contained in a disposable cassette device. The purification method results in highly pure genomic DNA over a wide range of input sample quantities, including as low as 1000 starting cells, and it gives high DNA yields while not relying on chromatography adsorption or solvent precipitation at any step. The purified DNA is suitable for use in PCR sequencing and in RFLP analysis. The disposable sample cassette is designed to process twelve or more samples in parallel and the processing instrument holds up to several of these cassettes. The instrument will inexpensive to manufacture and it will occupy less then one square foot of laboratory bench space.
The isolation of the genomic DNA from blood, bacteria, and virus is a necessary starting point for molecular diagnosis of infection, genetic disease, inherited traits and identity determination, as well as in research applications. The ability to rapidly and reproducibly isolate DNA from blood and other bodily samples is required to identify, characterize and treat factors involved in human disease and disorders.
DOE SBIR PHASE I GRANT #DE-FG-03-98ER82612
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