Nucleic Acids Research · 1997 · 3K citations · 8 references
Life SciencesMolecular BiologyMolecular GeneticsGenomicsPlant GeneticsPlant GenomicsNucleic Acid ExtractionNucleic Acid PurificationPolymerase Chain ReactionDna ExtractionGenomic Dna PurificationHigh-purity Nucleic Acid RecoveryPlant Nucleic Acid ExtractionRestriction Enzyme DigestionTissue Nucleic Acid ExtractionTotal Nucleic Acid Extraction
The study presents a simple, fast, universally applicable method for extracting high‑quality megabase genomic DNA from diverse organisms. The protocol uses inexpensive, non‑hazardous reagents, requires only 50–100 mg of tissue, and is suitable for low‑technology laboratories. The method successfully extracted high‑quality DNA from wheat, barley, potato, beans, pear, almond leaves, fungi, insects, and shrimp, enabling hundreds of PCR reactions and other downstream applications.
A very simple, fast, universally applicable and reproducible method to extract high quality megabase genomic DNA from different organisms is decribed. We applied the same method to extract high quality complex genomic DNA from different tissues (wheat, barley, potato, beans, pear and almond leaves as well as fungi, insects and shrimps' fresh tissue) without any modification. The method does not require expensive and environmentally hazardous reagents and equipment. It can be performed even in low technology laboratories. The amount of tissue required by this method is ∼50–100 mg. The quantity and the quality of the DNA extracted by this method is high enough to perform hundreds of PCR-based reactions and also to be used in other DNA manipulation techniques such as restriction digestion, Southern blot and cloning.
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DNA polymorphisms amplified by arbitrary primers are useful as genetic markers
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Rapid isolation of high molecular weight plant DNA
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Fingerprinting genomes using PCR with arbitrary primers
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