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Current protocols in plant biology 2016-May

Genomic DNA Isolation from Maize (Zea mays) Leaves Using a Simple, High-Throughput Protocol.

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Kristen Leach
Paula McSteen
David Braun

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Abstrak

A simple, robust, inexpensive, high-throughput method for isolating genomic DNA from maize (Zea mays) leaf tissues is described. The DNA obtained using this extraction protocol is suitable for polymerase chain reaction (PCR) genotyping, which can be employed for the identification of alleles in diverse genetic and breeding approaches, such as marker-assisted selection, genetic fine mapping, and mutant introgression. This method utilizes 96-well plates for the collection of leaf tissue and the subsequent isolation of genomic DNA. The DNA isolation step is performed inexpensively within 3 hr and uses a urea-based extraction buffer that does not require an organic extraction step. Yields of genomic DNA are sufficient to perform ∼25 PCR-genotyping reactions per sample. These qualities, coupled with the protocol being robust and easy for inexperienced users to master, make this method ideal for new researchers. © 2016 by John Wiley & Sons, Inc.

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