Heterodera glycines: density-based gradient separation of eggs.
An efficient technique was developed for separating early and late stages of embryonic development in eggs of Heterodera glycines. This technique takes advantage of density changes that occur during embryogenesis in the developing embryo and egg to partition the egg within a sucrose step gradient. Sorted samples of eggs separated with 82% enrichment for pre-gastrula early embryos and 93% enrichment for first and second stage unhatched juveniles as late embryos. Subpopulations enriched for either developmental stage are available for use in generating stage-specific cDNA libraries, normalization of subpopulations to synchronize development, biochemical characterization, and many other uses.
Biology Department, Saint Louis University, 3507 Laclede Ave, St Louis, MO 63103, USA.
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Burgwyn
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Cook
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Bolla
D000818:Animals; D029742:Caenorhabditis elegans Proteins / Q000096:biosynthesis / Q000235:genetics; D002499:Centrifugation, Density Gradient / Q000379:methods; D018076:DNA, Complementary / Q000096:biosynthesis; D005260:Female; D018507:Gene Expression Regulation, Developmental; D015801:Helminth Proteins / Q000096:biosynthesis / Q000235:genetics; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000235:genetics; D010063:Ovum; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013025:Glycine max / Q000469:parasitology*; D014431:Tylenchoidea / Q000196:embryology* / Q000235:genetics / Q000302:isolation & purification*
D029742:Caenorhabditis elegans Proteins; D018076:DNA, Complementary; D015801:Helminth Proteins; D018510:RNA, Helminth; C102163:HCH-1 protein, C elegans; D008666:Metalloendopeptidases