Preparation of Rice Plant Genomic DNA for Various Applications.

Research on plant molecularbiology, genetic engineering, and crop molecular breeding involves various manipulations of plant genomic DNAs (gDNAs). These studies require preparing gDNAs from plant materials using suitable methods according to the yield, intactness, and purity of the resultant gDNA samples.

Here we describe protocols for preparation of rice plant gDNAs for various applications including:

(1) maxipreps and

(2) minipreps of gDNAs for restriction analysis, gene cloning, PCR amplification, genomic sequencing, and genotyping;

(3) preparation of megabase-sized nuclear DNA for construction of large-insert genomic libraries and long-range physical mapping; and

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Construction of a quencher-free cascade amplification system for highly specific and sensitive detection of serum circulating miRNAs.

(4) 96-well-format high-throughput gDNA preparation for PCR-based genotyping. The methods are also suitable for other plant species including dicots.