ISSN: 0041-3216 (Online), 0041-3216 (Print)
Volume 87 Number 2
An improved protocol for isolation of high quality DNA from spathe and leaf tissue of Anthurium andraeanum (Hort.). (65)
DNA isolation was attempted using three protocols with two tissues (leaf vs. spathe) in two cultivars (Venus vs. Success) of Anthurium andraeanum (Hort.) with the aim of developing a robust, repeatable protocol capable of producing DNA of high quality and quantity. The three protocols were the Dellaporta protocol of 1983, a modified Rowhani protocol recommended by Buldewo and Jaufeerally-Fakim, and a modified Kobayashi (1998) protocol. The experiment was replicated three times with two samples per replicate and the entire trial repeated once again. The results indicated that although all the three protocols were capable of giving good yields (19-27 micro g g-1), only the Dellaporta protocol gave undegraded DNA of high integrity. Among the tissue types the leaf tissue (24.7 mocro g g-1) gave a much higher DNA yield than spathe tissue (19.9 micro g g-1). All protocols gave DNA that was PCR amplifiable and capable of digestion with the restriction enzyme, BamH1. Furthermore all the protocols gave acceptable purity levels estimated by OD 260/280 and OD 260/230. The results show that the Dellaporta protocol is the protocol of choice for library construction and other DNA manipulations.
Keywords: DNA purity, restriction analysis, DNA yield