Tb green rt pcr kit

The W303α strain was grown at 30°C until the OD₆₀₀ reached to 0.5. It was subsequently exposed to 0.15% MMS for 2 h. Total RNA was isolated from the untreated and MMS-treated cells using the acid phenol method as described previously (Suhane et al., 2014 (link)). Further cDNA was synthesized using reverse transcriptase (Omni Script; Qiagen, Hilden, Germany) as described previously (Suhane et al., 2014 (link)). The primer pair, OSB 14 and OSB 16, was used to amplify 307 bp at the 3′ end of ACT1 transcript. To amplify 326 bp at the 3′ end of RAD51, the primer pair OSB 44 and OSB 45 was used. Similarly, to amplify 291 bp at the 3′ end of the AHA1 transcript, the primer pair OSB 216 and OSB 394 was used. Finally, to amplify 276 bp at the 3′ end of SBA1, the primer pair OSB 391 and OSB 445 was used. For real-time RT-PCR, cDNA was diluted (1:50) and used for PCR using a Takara TB Green RT-PCR kit as described earlier (Suhane et al., 2014 (link)). The mean values (±SD) from three independent experiments were plotted using GraphPad Prism 6 software.