OPTIMIZING GENOMIC DNA ISOLATION IN PINEAPPLE (ANANAS COMOSUS L.)

Achigan-Dako E. Gbènato, Agbangla Clément

Abstract


The isolation of high molecular mass genomic DNA (Deoxyribonucleic acid) is crucial for applications in molecular biology. To this end many protocols were developed for the extraction of plant DNA. However, for pineapple (Ananas comosus), standard protocols are scarce and not always efficient when resources are limited. In this study, we developed a new protocol for nuclear DNA extraction in pineapple. Four existing protocols were tested but none has provided high quality DNA extract. The original laboratory standard protocol based on the use of CTAB (Cetyltrimethylammonium bromide) was successfully modified to optimize the quality of the DNA extract using eighteen pineapple young leaf samples including three parts: the leaf apex, the mid blade, and the leaf base of two cultivars (i.e. Sugarloaf and Smooth Cayenne). The successful extraction of DNA in a sister species, Ananas bracteatus, gave evidence that the protocol can be used for others Bromeliaceae. The new protocol yielded 51.76 µg/ml DNA, which is higher than that obtained with previous protocols. The DNA extract was efficiently PCR (Polymerase Chain Reaction) amplified using simple sequence repeat primers. We proposed henceforth the use of this protocol for further DNA isolation in pineapple particularly under limited resources condition when using CTAB.


Keywords


Ananas bracteatus; Ananas comosus; Bromeliaceae; CTAB; genomic DNA extraction; MATAB

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References


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