DNA microsatellites for the development of reliable PCR-based length polymorphisms in Phaseolus

By:

González, Delkin Orlando

Contributor(s):

Material type: Article

ArticleLanguage: English Description: p. 76-80Subject(s): LOC classification:

SB 327 .P5

In: In: Roca, William M.; Mayer, Jorge Edgard; Pastor Corrales, Marcial Antonio; Tohme M., Joseph (eds.). International Scientific Meeting Phaseolus Beans Advanced Biotechnology Research Network (2, 1993, Cali, Colombia). Phaseolus Beans Advanced Biotechnology Research Network: ProceedingsSummary: DNA microsatellites or Simple Sequence Repeats (SSR), consist of very short DNA sequences of repeated nucleotide motifs. Microsatellites were found to be abundant in human, mouse and more recently in rice, tomato and soybean. They have been shown to detect a high level of polymorphism making them ideal in the fine mapping of tagged genes, the detection of polymorphism in intraspecific crosses and population genetic studies. We have initiated a project to assess the presence, abundance of microsatellites in common bean and their potential as markers in mapping. A bean DNA genomic library was constructed by digesting total DNA to completion with Sau 3AI and cloning the fragments of less than 500 bp into the Xho I site of Lambda ZAP II. The library was screened with synthetic oligonucleotides, composed of di, tri or tetranucleotides repeats 8-18 times. The oligonucleotides were selected from previous reports on plant microsatellites or designed for this study. A first screening of the library using these oligonucleotides suggests that similar to other plant species CA repeats are present at very low frequency in bean in constrast with the human genome. (ATT)n and (AT)n repeats are so far the most abundant. Work is in progress to purify positive clones, sequence them, and design specific PCR primers flanking the microsatellites for use as new molecular markers in bean mapping.

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Item type	Current library	Collection	Call number	Copy number	Status	Date due	Barcode	Item holds
Books	CIAT Library CIAT Publications	CIAT Publications	SB 327 .P5 (Browse shelf(Opens below))		Available
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DNA microsatellites or Simple Sequence Repeats (SSR), consist of very short DNA sequences of repeated nucleotide motifs. Microsatellites were found to be abundant in human, mouse and more recently in rice, tomato and soybean. They have been shown to detect a high level of polymorphism making them ideal in the fine mapping of tagged genes, the detection of polymorphism in intraspecific crosses and population genetic studies. We have initiated a project to assess the presence, abundance of microsatellites in common bean and their potential as markers in mapping. A bean DNA genomic library was constructed by digesting total DNA to completion with Sau 3AI and cloning the fragments of less than 500 bp into the Xho I site of Lambda ZAP II. The library was screened with synthetic oligonucleotides, composed of di, tri or tetranucleotides repeats 8-18 times. The oligonucleotides were selected from previous reports on plant microsatellites or designed for this study. A first screening of the library using these oligonucleotides suggests that similar to other plant species CA repeats are present at very low frequency in bean in constrast with the human genome. (ATT)n and (AT)n repeats are so far the most abundant. Work is in progress to purify positive clones, sequence them, and design specific PCR primers flanking the microsatellites for use as new molecular markers in bean mapping. eng