Chromatography in DNA radiolabeling: Hands-off automation using a robotic workstation |
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| Institution: | 1. Department of Biochemistry and Molecular Biology, Ferrara University, Via L. Borsari 46, 44100 Ferrara, Italy;2. Beckman Analytical, Via Cintia 41/F, 80126 Naples, Italy;3. Biotechnology Center, Ferrara University, Via Fossato di Mortara 64, 44100 Ferrara, Italy;1. School of Chemistry and Chemical Engineering, Shandong University, 250100, Jinan, PR China;2. Breast Center, Central Hospital Affiliated to Shandong First Medical University, 250013, Jinan, PR China;3. Shandong Academy of Preventive Medicine, Shandong Center for Food Safety Risk Assessment, Shandong Center for Disease Control and Prevention, 250014, Jinan, PR China;1. School of Chemistry and Chemical Engineering, Chongqing University of Technology, Chongqing, 400054, PR China;2. Key Laboratory of Luminescence Analysis and Molecular Sensing, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing, 400715, PR China;1. College of Life Sciences, Jilin Agricultural University, Changchun 130118, China;2. Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun 130122, China |
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| Abstract: | The experiments described in the present paper were performed in order to determine whether the Biomek-1000 (Beckman Instruments, Fullerton, CA, USA) automated laboratory workstation can be used in a fully automated DNA labeling method followed by automated gravity-driven size exclusion purification of molecular probes. To this aim, we performed random oligodeoxyribonucleotide priming of a HIV-1 LTR probe that was used for molecular hybridization to Southern blotted polymerase chain reaction products. The results obtained demonstrate that the automatically labeled probe can be efficiently purified by automated and gravity-driven Sephadex G-50 chromatography, without any major changes in hybridization property. This robotic methodology can be used in several procedures employing radioisotope labeling. |
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