Optimization of genomic DNA isolation from Cupressus sempervirens L. and development of RAPD markers for breeding applications
1997
Ghosn, D.
Efficient total genomic DNA isolation, compatible with cloning or PCR requirements, is a first step towards molecular studies of a species. Optimisation of a method for reproducibly obtaining high quality DNA is particularly important in research involving fingerprinting of recalcitrant forest species grown in natural stands or in provenance tests in the field. We describe herein the adaption of a modified CTAB procedure to isolate DNA from the cypress (Cupressus sempervirens L.), a typical conifer of the Mediterranean region, and some preliminary data on PCR based provenance fingerprinting. The extraction method employed, yielded DNA of higher quantity and purity when compared with SDS phenolchlorophorm protocols. A series of RAPD PCR optimisation experiments were conducted in order to develop a protocol adapted to cypress and of sufficient reproducibility.
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