Abstract
ABSTRACT The application of molecular techniques to tackle ecological and evolutionary questions requires genomic DNA in good quality and quantity. The quality of the isolated DNA, however, can be influenced by the tissue type and the way the sample was conserved and manipulated. Therefore, customizing protocols to improve the DNA isolation and locus amplification is crucial. We optimized a cheap and manual protocol of DNA extraction and microsatellites amplification using five different tissues of a palm species of the brazilian Atlantic Forest. We successfully extracted DNA from all five tissue types. Leaf, stem, and endocarp of non-dispersed seeds presented the highest rates of successful DNA extraction and microsatellite amplification; whereas root, endocarp of dispersed seeds, and embryo showed the lowest quality and quantity. Based on these results, we discussed the implications of using different tissues for studies about seed dispersal, pollination, and population genetics.
Highlights
Molecular tools have been used to answer different ecological and evolutionary questions, and the isolation of DNA in good quantity and quality is a critical step towards the development of these studies (Haig, 1998, Allen et al 2006)
We report a DNA amplification protocol using microsatellite loci, in which we obtained high quality results even using low amounts of DNA template
DNA isolation from roots and embryos was less successful and microsatellite amplification was lower for DNA extracted from roots and endocarps of dispersed seeds
Summary
Molecular tools have been used to answer different ecological and evolutionary questions, and the isolation of DNA in good quantity and quality is a critical step towards the development of these studies (Haig, 1998, Allen et al 2006). DNA extraction from plants is inherently more difficult than from animals, first because plants have cell wall and lignin (Varner & Lin 1989) that require maceration steps to be broken, and second because plants often contain secondary compounds that can inhibit PCR (Khanuja et al 1999). DNA can be extracted from various tissues, such as leaf, stem, root, fruit endocarp and embryo. These tissues can be used to answer different ecological and microevolutionary questions. For example, are often used in population genetics
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