Mitochondrial rRNA Oligonucleotide Synthesis

In this study it was used the gene encoding the mitochondrial large-subunit of P. jirovecii (mt LSU rRNA). Design of oligonucleotides and in silico analysis is described in the Electronic Supplementary Information (Scheme S1). Table 1 shows the sequences of the oligonucleotides employed in this study. The synthesis took place by the well-defined phosphoramidite solid phase protocol [32 (link)]. For this, oligonucleotides were assembled on controlled pore glass (CPG) scaffolds by consecutive incorporation of the suitable phosphoramidites employing an automated Applied Biosystems 394 DNA synthesizer (Foster City, CA, USA). Then, the scaffolds were treated overnight with concentrated ammonia at 55 °C. Finally, synthetized oligonucleotides were purified by Glen-PackTM DNA cartridges (Glen Research, Sterling, VA, USA) and mass spectrometry was used to analyze them (Table S1, ESI).