Isolation of genomic DNAs from the tropical fruit trees avocado, coconut, guava and mango for PCR-based DNA marker application
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Abstract
DNA with sufficient quality for the application of PCR-based
DNA marker technology very often has severe problems due
to the presence of inhibitors such as polysaccharides, which
inhibit enzymatic DNA processing or polyphenols as inhibitors
of PCR reactions. Here, different protocols for DNA extraction
and purification were tested with the four tropical fruit trees
guava (Psidium guajava L.), avocado (Persea americana
Mill.), mango (Mangifera indica L.) and coconut (Cocos
nucifera L.). The well-established CTAB protocol of Doyle
and Doyle yielded excellent DNA templates for PCR
amplification with mango and coconut, but not so with guava
and avocado. For these latter species, several other extraction
protocols also yielded only non-satisfactory results.
Modification of the CTAB method with respect to CTAB buffer
composition and in combination with reversible adsorption
to NucleoSpin columns alleviated the problems encountered
with the genomic DNA of both species. The quality of DNA
prepared by this procedure allowed AFLP, SSR and ISTR DNA
marker analyses in guava and/or avocado.
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