Serial extraction of endosperm drillings (SEED)—A method for detecting transgenes and proteins in single viable maize kernels

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2001-06-01
Authors
Sangtong, Varaporn
Mottl, Erik
Long, Mary Jane
Lee, Michael
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Scott, M. Paul
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Agronomy

The Department of Agronomy seeks to teach the study of the farm-field, its crops, and its science and management. It originally consisted of three sub-departments to do this: Soils, Farm-Crops, and Agricultural Engineering (which became its own department in 1907). Today, the department teaches crop sciences and breeding, soil sciences, meteorology, agroecology, and biotechnology.

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The Department of Agronomy was formed in 1902. From 1917 to 1935 it was known as the Department of Farm Crops and Soils.

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1902–present

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  • Department of Farm Crops and Soils (1917–1935)

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Abstract

We have developed a method for detecting a transgene and its protein product in maize endosperm that allows the kernel to be germinated after analysis. This technique could be highly useful for several monocots and dicots. Our method involves first sampling the endosperm with a hand-held rotary grinder so that the embryo is preserved and capable of germination. This tissue is then serially extracted, first with SDS-PAGE sample buffer to extract proteins, then with an aqueous buffer to extract DNA. The product of the transgene can be detected in the first extract by SDS-PAGE with visualization by total protein staining or immuno-blot detection. The second extract can be purified and used as template DNA in PCR reactions to detect the transgene. This method is particularly useful for screening transgenic kernels in breeding experiments and testing for gene silencing in kernels.

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This article is published as Sangtong, Varaporn, Erik C. Mottl, Mary Jane Long, Michael Lee, and M. Paul Scott. "Serial extraction of endosperm drillings (seed)—A method for detecting transgenes and proteins in single viable maize kernels." Plant Molecular Biology Reporter 19, no. 2 (2001): 151-158.

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