Separation of integument and nucellar tissues from cotton ovules (Gossypium hirsutum L.) for both high- and low-throughput molecular applications
2013
Frank Bedon | Lisa Ziolkowski | Kenji Osabe | Ingrid Venables | Adriane Machado | Danny Llewellyn
Here we present a quick and low-cost method to separate the different layers of tissue from the ovules and young seeds of cotton (Gossypium hirsutum L.) for use in high- and low-throughput molecular applications. This method is performed at room temperature using standard laboratory equipment and does not require embedding of the samples, time-consuming fixation, or micro-sectioning procedures. We show that the three main tissues can be efficiently separated from isolated ovules collected on the day of anthesis. RNA and genomic DNA extracted from tissues separated by this method are of good quality and suitable for a variety of molecular applications to study the early stages of cotton seed and fiber development.
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