High-Cot sequence analysis of the maize genome.
Higher eukaryotic genomes, including those from plants, contain large amounts of repetitive DNA that complicate genome analysis. We have developed a technique based on DNA renaturation which normalizes repetitive DNA, and thereby allows a more efficient outcome for full genome shotgun sequencing. The data indicate that sequencing the unrenatured outcome of a Cot experiment, otherwise known as High-Cot DNA, enriches genic sequences by more than fourfold in maize, from 5% for a random library to more than 20% for a High-Cot library. Using this approach, we predict that gene discovery would be greater than 95% and that the number of sequencing runs required to sequence the full gene space in maize would be at least fourfold lower than that required for full-genome shotgun sequencing.
Yuan
Yinan
Y
Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907-1392, USA.
SanMiguel
Phillip J
PJ
Bennetzen
Jeffrey L
JL
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D018745:Genome, Plant; D015698:Genomic Library; D020071:Interspersed Repetitive Sequences / Q000235:genetics; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D012689:Sequence Homology, Nucleic Acid; D003313:Zea mays / Q000235:genetics*