Amplification of 5′RLM-RACE products was performed (1× High Fidelity PCR buffer, 0.6 µM GeneRacer 5′ primer, 0.2 µM of the gene specific primer (electronic supplementary material, table S1), 200 µM dNTPs, 1 mM MgSO4, 3% DMSO and 0.5U Platinum Taq DNA Polymerase High Fidelity) followed by a nested PCR, using 1 µl of the PCR product in a 50 µl reaction (1× High Fidelity PCR buffer, 0.2 µM GeneRacer 5′ nested primer, 0.2 µM of the nested gene specific primer (electronic supplementary material, table S1), 200 µM dNTPs, 1 mM MgSO4 and 0.5U Platinum Taq DNA Polymerase High Fidelity).
PCR products were amplified with a Phusion High-Fidelity DNA Polymerase (New England Biolabs, Massachusetts, USA) and cloned using a Zero Blunt TOPO PCR Cloning Kit (Invitrogen, California, USA).
de Vries S., Kukuk A., von Dahlen J.K., Schnake A., Kloesges T, & Rose L.E. (2018). Expression profiling across wild and cultivated tomatoes supports the relevance of early miR482/2118 suppression for Phytophthora resistance. Proceedings of the Royal Society B: Biological Sciences, 285(1873), 20172560.
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