RNA-Seq of Ralstonia syzygii subsp. celebesensis
Genomic DNA was extracted from Ralstonia syzygii subsp. celebesensis strain Bbd-F-1 using the Quick-DNA Magbead Plus Kit (Zymo Research, D4082). DNA quality was verified via Nanodrop (251.7 ng/L, A260/280=1.93). PCR Amplification was using Primers: 121F/121R (Ray et al 2021), Mix: MyTaq HS Red Mix (Bioline, BIO-25048), Cycling: 95C (2 min); 34 cycles of 95C (30 s), 53C (20 s), 72C (1 min); final extension at 72C (5 min), Instrument: VeritiPro Thermal Cycler (Applied Biosystems). Sequencing method was using Bidirectional Sanger sequencing (capillary electrophoresis), on Platform: Biosystems 3500xl. The output was a 317 bp consensus sequence that was confirmed by BLAST against Ralstonia NCBI entries.