Illumina MiSeq paired end sequencing; The effect of the pBCXO1 plasmid on the transcriptome of Bacillus cereus G9241 at both 37  and 25 degree celsius
The effect of the pBCXO1 plasmid on the transcriptome of Bacillus cereus G9241 at both 37  and 25 degree celsius
Bacterial cultures were grown to exponential phase at 25 or 37˚ degree celsius before samples were taken and RNAprotect (Qiagen) was added to each samples. QIAzol (Qiagen) was added to each pelleted samples before transferring to Lysing Matrix B tubes (MP Biomedicals). Cells were lysed using the FastPrep®-24 Classic instrument (MP Biomedicals) with a COOLPREP™ adapter (MP Biomedicals). Bead beating was conducted at 6 ms-1 for 40 s for 2 cycles, with a 300 s pause between cycles. Lysates were centrifuged for 1 minute at 10,000 g and supernatant extracted. an RNeasy Micro Kit (Qiagen) was then used to extract RNA following manufacturers protocol. TruSeq library prep was conducted using the Low Sample (LS) protocol and manufacturer's instructions were followed which are detailed in the LS protocol of the TruSeq® Stranded mRNA Sample Preparation Guide (Part # 15031047 Rev. E).