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United States Department of Agriculture

Agricultural Research Service

Title: High throughput tissue preparation for large-scale genotyping experiments

Authors
item Boda Slotta, Tracey - USDA-ARS, FARGO, ND
item Brady, Lee - USDA-ARS, FARGO, ND
item CHAO, SHIAOMAN

Submitted to: Molecular Ecology Notes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: June 11, 2007
Publication Date: January 1, 2008
Repository URL: http://hdl.handle.net/10113/32698
Citation: Boda Slotta, T.A., Brady, L., Chao, S. 2008. High throughput tissue preparation for large-scale genotyping experiments. Molecular Ecology Resources. 8:83-87

Interpretive Summary: The manuscript presents a protocol for sample preparation for DNA extraction on a large-scale. Tissue samples were collected in the presence of silica gel desiccant in order to dry and preserve the tissue prior to DNA extraction. For large scale sample preparation, a 96-well plate format is used with a fixed amount of silica gel in each well. This allows users to collect 96 samples in each batch by inserting sample tissues into each well. Samples prepared in this manner can be collected by collaborators and sent through the mail (without ice or cold packs) to the lab for further sample processing. Prior to DNA extraction, silica gel can be further used to aid grinding of sample tissues into powder. For the plant and animal tissue included in the study, the quantity of silica gel used was not as important as the quality of the tissue collected. However, inclusion of too much tissue can prevent samples from drying and creates a risk of sample deterioration.

Technical Abstract: A fast, efficient technique is described for the extraction of DNA from a large number of samples. The applications of this method include population genetics, plant breeding, and genetic screening. In the field, samples are collected in premeasured silica gel aliquots in polypropylene blocks, which are later used to grind the dried tissue. This permits naturalists, breeders, and collaborators to collect a large number of samples in a short amount of time and allows the samples to dry quickly during shipping. No phenol or chloroform steps are required to obtain high-quality DNA. Samples representing 12 plant families, three invertebrates, and a mammal were included. Quantities of DNA obtained were consistent with or better than other techniques. The quality of samples was tested by amplification of the internal transcribed spacer region. Test amplifications were successful, confirming the quality of extracted DNA.

Last Modified: 9/10/2014
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