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United States Department of Agriculture

Agricultural Research Service

Title: USING PSTL AS A METHOD OF GENOME FILTERING IN WHEAT

Author
item Fellers, John

Submitted to: Plant Genome Conference Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: October 18, 2004
Publication Date: January 10, 2005
Citation: Fellers, J.P. 2005. Using pstl as a method of genome filtering in wheat. Plant Genome Conference Proceedings.

Technical Abstract: Bread wheat contains one of the largest genomes of the plant kingdom and is a significant challenge when considered for genome sequencing. Wheat is known to have gene-rich regions within the genome interspersed by vast amounts of repetitive DNA. It is understood that a filtering technique would be of significant benefit when targeting gene-containing regions for sequencing. Previous work has indicated that the restriction enzyme, Pst1, can increase the presence of low copy DNA when used to digest genomic DNA. To evaluate this on a larger scale, genomic DNA of the bread wheat "Chinese Spring" was digested with the methyl sensitive Pst1. DNA fragments of 2-4 Kb were subcloned into cloning vector, isolated, and single pass sequenced. The project goal is to isolate and sequence more than 3,000 clones to get a sub sample of the capability of this technique. The sequencing results will be evaluated for the presence of open reading frames, repetitive DNA and low copy DNA.

Last Modified: 7/10/2014
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