ACRI_CL78 Tandem Repeat Probe Labeling

A probe for A. cristatum tandem repeat ACRI_CL78 (Said et al. 2018 (link)) was labeled by PCR with digoxigenin-dUTP (Roche, Mannheim, Germany) using diploid A. cristatum cv. Parkway DNA as a template. Biotin-dUTP (Roche) or digoxigenin-dUTP labeled probe for 5S rDNA was prepared according to Fukui et al. (1994 (link)) using rice DNA as a template for PCR. The plasmid pTa71 (45S rDNA) containing a 9-kb fragment from bread wheat with 18S-5.8S-26S rDNA and intergenic spacers (Gerlach and Bedbrook 1979 (link)) and genomic DNA of tetraploid A. cristatum PI 222957 were labeled with either biotin or digoxigenin by nick translation using standard kits (Nick Translation Mix, Roche) following the manufacturer’s instructions.

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Said M., Parada A.C., Gaál E., Molnár I., Cabrera A., Doležel J, & Vrána J. (2019). Uncovering homeologous relationships between tetraploid Agropyron cristatum and bread wheat genomes using COS markers. TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik, 132(10), 2881-2898.

Publication 2019

digoxigenin-dUTP Biotin-dUTP Nick Translation Mix

Biotin Bread Digoxigenin Diploid Dutp Genomic Plasmid Rdna Rice Tandem repeat Tetraploid Wheat

Corresponding Organization :

Other organizations : Czech Academy of Sciences, Institute of Experimental Botany, University of Córdoba, Agricultural Institute, Centre for Agricultural Research, Hungarian Academy of Sciences

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Variable analysis

independent variables

Probe labeling method (PCR with digoxigenin-dUTP or biotin-dUTP)

dependent variables

Hybridization of labeled probes

control variables

Diploid A. cristatum cv. Parkway DNA as template for ACRI_CL78 probe
Rice DNA as template for 5S rDNA probe
Plasmid pTa71 (45S rDNA) containing a 9-kb fragment from bread wheat
Genomic DNA of tetraploid A. cristatum PI 222957

controls

Positive control: Diploid A. cristatum cv. Parkway DNA as template for ACRI_CL78 probe
Positive control: Rice DNA as template for 5S rDNA probe
Positive control: Plasmid pTa71 (45S rDNA) containing a 9-kb fragment from bread wheat
Positive control: Genomic DNA of tetraploid A. cristatum PI 222957

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