Nuclear isolation and purification using SDS/urea (NIPSU) method for efficient and rapid extraction of high-purity genomic DNAs from Jatropha curcas L: A comparative analysis of DNA isolation protocols

摘要

Jatropha curcas L. (Jatropha) has gained popularity for its potential use in biodiesel production in arid regions, and its yield improvement by molecular breeding has been anticipated. However, Jatropha is known for its recalcitrance in the extraction of pure genomic DNAs. In this study, four DNA extraction methods were comparatively examined for their efficiency and quality of genomic DNA isolations from Jatropha. Consequently, one method, designated as nuclear isolation and purification using SDS/urea (NIPSU) method was found to be the most effective for rapid preparation of high yield and quality genomic DNA from Jatropha leaves. NIPSU method allowed extraction of genomic DNAs from leaves, stems, and roots tissues of Jatropha in high yield (6.8 to 24.5 μg gFW-1), with spectroscopically high purity (A260/A280 and A260/A230 >1.8). NIPSU method was also applicable to the dehydrated leaves for genomic DNA preparations. The isolated genomic DNA was amenable for downstream molecular procedures such as polymerase chain reaction, restriction enzyme cutting and Southern blotting analyses.