Isolation of megabase-size DNA from sorghum and applications for physical mapping and bacterial and yeast artificial chromosome library construction |
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| Authors: | Sung-Sick Woo Vipin K. Rastogi Hong-Bin Zhang Andrew H. Paterson Keith F. Schertz Rod A. Wing |
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| Affiliation: | (1) Department of Soil and Crop Sciences, Texas A&M University, 77843 College Station, TX, USA;(2) USDA-ARS, Texas A&M University, 77843 TX, College Station, USA;(3) Present address: Department of Biochemistry and Biophysics, Texas A&M University, USA |
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| Abstract: | A method was developed for the isolation of megabase-size DNA fromSorghum bicolor. Sorghum protoplasts were isolated from young leaf tissue, embedded in an agarose matrix as microbeads or plugs, followed by cell lysis and protein degradation. The DNA prepared by this method was larger than 1 Mb in size and readily digestible with restriction enzymes. The DNA was shown to be suitable for physical mapping, and was successfully used for the construction of BAC and YAC libraries. |
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| Keywords: | Sorghum bicolor Megabase-size DNA YAC and BAC cloning Physical mapping |
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