title: Excised radicle tips as a source of genomic DNA for PCR-based genotyping and melting curve analysis in cotton.
abstract: Genomic DNA isolation in cotton is complicated because of the presence of secondary metabolites that are inhibitory to PCR amplification. We report here that radicle tips, but not other parts of cotton seedlings, yield high-quality DNA that is readily amenable for PCR. The radicle-tip-excised seedlings retain viability because of the formation of adventitious roots. We demonstrate the utility of this method in distinguishing homozygotes from heterozygotes in a cotton breeding population and in hybrid seed purity testing.
authors: Centre for Cellular and Molecular Biology (CCMB), Uppal Road, Hyderabad 500 007, India.
authors: P Srinivasa
authors: PS
authors: Rao
authors: P Sateesh
authors: PS
authors: Kumar
authors: Ramesh V
authors: RV
authors: Sonti
mesh_terms: D002678:Chimera / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification*; D005838:Genotype; D060005:Genotyping Techniques; D018525:Germination; D003368:Gossypium / Q000235:genetics*; D006579:Heterozygote; D006720:Homozygote; D009691:Nucleic Acid Denaturation; D018517:Plant Roots / Q000235:genetics*; D016133:Polymerase Chain Reaction; D036226:Seedlings / Q000235:genetics*
chemical_list: D018744:DNA, Plant
references: New Phytol. 2008;180(3):594-607
references: 18684160
references: Clin Chem. 2003 Jun;49(6 Pt 1):853-60
references: 12765979
references: Hum Mutat. 2009 Jun;30(6):860-6
references: 19418555
references: Am J Pathol. 1998 Oct;153(4):1055-61
references: 9777937
references: Anal Biochem. 1997 Feb 15;245(2):154-60
references: 9056205
references: Genomics. 2008 Nov;92(5):353-8
references: 18721872
references: BMC Genet. 2008 Nov 03;9:69
references: 18980665
references: Methods Enzymol. 1974;31:528-44
references: 4418408
references: Theor Appl Genet. 2008 Sep;117(5):721-8
references: 18553067
references: Anal Biochem. 1993 Apr;210(1):34-8
references: 8098189