DOE Human Genome Program Contractor-Grantee Workshop VIII
February 27-March 2, 2000  Santa Fe, NM

44. Development of a High Throughput Peptide Nucleic Acid Synthesizer

J. Shawn Roach¹, Simon Rayner¹, Lynn Mayfield², David R. Corey², and Harold "Skip" Garner¹

¹Center for Biomedical Inventions, Department of Internal Medicine and ²Howard Hughes Medical Institute, Department of Pharmacology and Department of Biochemistry, University of Texas Southwestern Medical Center at Dallas, Dallas, TX

roach@ryburn.swmed.edu

Peptide Nucleic Acids (PNAs) are synthetic analogs of DNA in which the phosphodiester backbone has been replaced with 2-aminoethyl glycine linkages, but maintaining the four natural nucleobases. A PNA strand will bind to DNA with the same sequence complementarity as standard DNA/DNA base paring, but PNA/DNA binding occurs more rapidly and more tightly than DNA/DNA binding. Much research has gone into the potential applications of PNAs as antisense and diagnostic agents.¹ However, a major obstacle in PNA research becoming more widespread has been the high cost of the PNAs. A high throughput PNA synthesizer will afford an economy of scale and reduce the synthetic cost of PNAs. We report the development of a high throughput PNA synthesizer capable of producing up to 192 different PNAs in one synthesis run. The synthesizer is based on high throughput DNA synthesis technology developed at the University of Texas Southwestern Medical Center at Dallas to support the Human Genome Project.² The synthesizer consists of an XY table, a series of valves plumbed to an injection head for reagent delivery, two vacuum chucks for reagent removal and a computer that controls the synthesis procedure. Synthesis is conducted in 96-well fritted filter plates, using standard solid phase Fmoc-PNA synthesis chemistry. The quality of the PNAs produced from the synthesizer is assessed using RP-HPLC and MALDI MS.

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