Optimization of genomic DNA extraction protocol for black wattle

As the integration of classic techniques and molecular genetics analysis is a current trend in forest tree improvement, a protocol for DNA extraction adapted for the study of each species is necessary. The objective of this study was to determine the effect of different adaptations in the CTAB protocol of genomic DNA extraction for black wattle. The following components were tested after the organic extraction phase: chloroform, phenol formaldehyde, and proteinase K. Also, the effects of applying RNase after the precipitation and cleaning phase were analyzed. The efficiency of these treatments was evaluated on fresh leaflets samples, as well as on samples refrigerated for seven days. The authors observed that all treatments enabled the successful genomic DNA extraction using the CTAB protocol. Moreover, storage did not affect the DNA integrity; adding RNase, however, resulted in an enhanced extracted DNA. Therefore, the use of chloroform and RNase according to the CTAB protocol on both fresh and refrigerated samples is suggested.