A Rapid Microwave Method for Isolation of Genomic DNA and Identification of White Rot Fungi

A rapid, effective and efficient method to identify the innumerable white rot fungal strains is of utmost importance. Mycelia of the unknown as well as know isolates of WRF, after alternative washing with TE buffer and sterile water, were suspended in TE buffer. Fungi in solution were then exposed to microwave. The crude extract contained genomic DNA which was extracted and amplified using ITS primers for further identification. Based on sequencing results the identity of known cultures was confirmed, while the unknown cultures were identified as Clitopilus scyphoides (AGUM004, BankIt2098576 MH172163); Ganoderma rasinaceum (AGUM007, BankIt2098576 MH172163); Schizophyllum sp (KONA001 BankIt2098576 MH172164; AGUM011 BankIt2098576 MH172165 and AGUM021 BankIt2098576 MH172166 respectively), Coprinellus disseminatus (BANG001, BankIt2098576 MH172167) and Lentinus squarrosulus (TAMI004, BankIt2098576 MH172167). The microwave method described for isolating quality DNA of WRF without further purification steps proved a novel method requiring less than ten minutes and minimized the chances of the presence of PCR inhibitors.

Excised radicle tips as a source of genomic DNA for PCR-based genotyping and melting curve analysis in cotton.

Genomic DNA isolation in cotton is complicated because of the presence of secondary metabolites that are inhibitory to PCR amplification. We report here that radicle tips, but not other parts of cotton seedlings, yield high-quality DNA that is readily amenable for PCR. The radicle-tip-excised seedlings retain viability because of the formation of adventitious roots. We demonstrate the utility of this method in distinguishing homozygotes from heterozygotes in a cotton breeding population and in hybrid seed purity testing.

Optimization of conus genomic DNA isolation methods / 中国海洋药物

Conopeptides (Conotoxin) derived from tropical marine gastropod, cone snail ( Conus ) venoms have been an useful tool agent in neuro investigation and new drug source. Recently, new method of Conotoxin genes cloning from Conus genomic DNA has been established. So to get Conus genomic DNAs rapidly is the basis for diverse Conotoxin gene isolation, gene bank construction and the medical source exploitation. In this report, different DNA extractions from different tissue and organs of six cone snail species were performed in order to optimize the DNA isolation methods. The optimized brief phenol SDS method was established which fit for cone snails genomic DNA extraction.