RNA-seq of an inducible whiA CRISPRi strain grown with 0 or 500 ng/mL anhydrotetracycline
This experiment was performed to transriptioally profile bacteria with a whiA conditional knockdown using CRISPR interference (CRISPRi). A conditional knockdown strain was treated with 0 or 500 ng/mL anhydrotetracycline (ATC) for 4 days, which turns on the CRISPRi machinery. RNA extraction was performed using Zymo RNA miniprep kit. Libraries were constructed using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs) using 150 ng of input RNA. Libraries were amplified using 8 cycles on the thermocycler. Post amplification libraries were size selected ay 250-450 bp in length using SparQ beads. Libraries were validated using Bioanalyzer high sensitivity DNA analysis.
This experiment was performed to transriptioally profile bacteria with a whiA conditional knockdown using CRISPR interference (CRISPRi). A conditional knockdown strain was treated with 0 or 500 ng/mL anhydrotetracycline (ATC) for 4 days, which turns on the CRISPRi machinery. RNA extraction was performed using Zymo RNA miniprep kit. Libraries were constructed using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs) using 150 ng of input RNA. Libraries were amplified using 8 cycles on the thermocycler. Post amplification libraries were size selected ay 250-450 bp in length using SparQ beads. Libraries were validated using Bioanalyzer high sensitivity DNA analysis.