In the search for those mostly still-elusive SNPs that are widely thought to predispose to complex, multigenic diseases such as schizophrenia and heart disease, one technical hurdle that slows down the search is the sheer amount of sequencing required to scrutinize DNA from many healthy and sick individuals. A paper in this month's Nature Biotechnology offers an engineering improvement towards increasing the throughput of this effort.
Researchers at Columbia University in New York, led by Jingyue Ju, report that they have developed a way of creating combinatorial fluorescence energy transfer (CFET) tags that enables them to run eight tests at once where before they could have only run one at a time. The method addresses the problem that, to date, only few fluorescent dyes with distinguishable energy spectra are available.
The researchers are exploiting combinatorial synthesis and energy transfer between the dyes. That increases the number of fluorescent signatures that can be read simultaneously even while exciting the tags with a single wavelength. The tags require a standard optical system containing three bandpass filters. In a test experiment, the scientists identified six nucleotide variations on a PCR product from a patient's retinoblastoma tumor suppressor gene
These CFTE tags can be a tool for high-throughput multicolor biological analysis of many problems, including multiplex immunoassays and detection of microdeletions and amplifications in chromosomes, the authors write.—Hakon Heimer
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- Tong AK, Li Z, Jones GS, Russo JJ, Ju J. Combinatorial fluorescence energy transfer tags for multiplex biological assays. Nat Biotechnol. 2001 Aug;19(8):756-9. PubMed.