Rneasy protect bacteria mini kit

Total bacterial RNA was extracted using the Qiagen RNeasy Protect Bacteria mini kit and treated with DNase I (Promega) and reverse transcription was performed using SuperScript III Reverse Transcriptase. The RT-qPCR assays were performed in the OneStepPlus Real-Time qPCR System (Applied Biosystems) using the Power SYBR Green PCR Master Mix (Applied Biosystems). The rtcB mRNA, rtcA mRNA, 23S rRNA, 16S rRNA and 5S rRNA sequences were amplified using the target-specific primer pairs 5′-ACG TGA TAA AGG TGC CTG GG-3′ and 5′-CAC ACC TGG TCC GAC TCA TC-3′; 5′-GAC CAA CTG GTG CTA CCG AT-3′ and 5′-GCG TTA CGC CAT CTG TTT CT-3′; 5′-AGA GTA ACG GAG GAG CAC GA-3′ and 5′-CAC TAT GAC CTG CTT TCG CA-3′; 5′-CGG ACG GGT GAG TAA TGT CT-3′ and 5′-CTC AGA CCA GCT AGG GAT CG-3′; 5′-GGT GGT CCC ACC TGA CCC-3′ and 5′-ATG CCT GGC AGT TCC CTA CT-3′, respectively. The 5S rRNA served as an endogenous control.

Total bacterial RNA was extracted using the Qiagen RNeasy Protect Bacteria mini kit and treated with DNase I (Promega) and reverse transcription was performed using SuperScript III Reverse Transcriptase. The RT-qPCR assays were performed in the OneStepPlus Real-Time qPCR System (Applied Biosystems) using the Power SYBR Green PCR Master Mix (Applied Biosystems). Selected genes were amplified using target-specific primer pairs as described previously.¹⁵ (link)