Comparative efficacy of different DNA extraction methods for PCR-based assay in Tectona grandis L.f.

Abstract

Four extraction methods and sample types were evaluated for yield, quality and suitability of genomic DNA for ISSR (inter simple sequence repeats) marker amplification by PCR in teak (Tectona grandis). Both CTAB (hexadecyltrimetylammonium bromide) and SDS (sodium dodecyl sulfate) procedures extracted large amounts of high purity genomic DNA. Leaves of trees, bud grafts and seedlings yielded better DNA than seeds. Using identical PCR conditions, DNA extraction methods and sample types affected amplifications of ISSR markers with a pattern: seedling > graft > tree leaves. Genomic DNA of seed samples contained high amount of protein and showed no ISSR amplifications, which were overcome by inclusion of proteinase K in SDS based extraction method. In conclusion, DNA extraction method and sample types are very important for reproducible ISSR-based molecular marker analysis in teak.