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Nucleic Acids Research, 1994, Vol. 22, No. 9 1701-1704
© 1994

METHODS

A rapid scanning strip for tri- and dinucleotide short tandem repeats

Manfred S. Wehnert1,4,*, Robert S. Matson3, Jang B. Rampal3, Peter J. Coassin3 and C.Thomas Caskey1,2

1Department of Molecular and Human Genetics 2Howard Hughes Medical Institute, Baylor College of Medicine Houston TX 77030 3Beckman Instruments, Inc., Advanced Development Unit Fullerton, CA 92634, USA 4Institute for Medical Genetics, University of Greifswald Greifswald, Germany

*To whom correspondence should be addressed at: Department of Molecular and Human Genetics, Baylor College of Medicine, Room T832, One Baylor Plaza, Houston, TX 77030, USA

Received December 16, 1993. Revised March 15, 1994. Accepted March 15, 1994.

Oligonucleotides representing 60 trinucleotide (21mers) and four dinucleotide (20mers) tandem repeats were directly synthesized and arrayed onto an aminated polypropylene substrate. DNA samples of different complexities (a CAG-containing 21mer oligonucleotide, PCR fragments of 200 to 3,000 bp, and cosmids with 31 to 35 kb inserts) were radiolabelled and hybridized to the oligonucleotide array at various temperatures. When compared to sequence data available from the test DNAs, the reverse blot system specifically identified various tri- and dinucleotide short tandem repeats (STRs) in every case. Moreover, there was no random or cross hybridization to nonspecific sequences. It was possible to detect as few as three repeated units in a particular location, as shown for (CCT)n, (GCC)n and (CAC)n triplets in cosmid DNA. Varying the hybridization stringency can enhance the detection of STRs. This single-step reverse blot system therefore allows the rapid, specific and sensitive identification of various STRs in DNA sources of different complexity.


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