CRISPRi_K12p_OriLyt_R_ALTp_Iytic_replicate_1
RNA from the CRISPRi and CRISPRa cells was purified by RNeasy Mini kit (Qiagen, #74106) according to the manufacturers instructions. The integrity of the isolated RNA was tested using the Agilent TS HSRNA Kit and Tapestation 4200 at the Genome Technology Center at the Azrieli Faculty of Medicine Bar-Ilan University. 1000 ng of total RNA were used for mRNA enrichment by using NEBNext mRNA polyA Isolation Module (NEB, #E7490L), and libraries for Illumina sequencing were performed using the NEBNext Ultra II RNA kit (NEB, #E7770L). Quantification of the library was performed using dsDNA HS Assay Kit and Qubit 2.0 (Molecular Probes, Life Technologies), and qualification was done using the Agilent D1000TS Kit and Tapestation 4200. 500 nM of each library was pooled together and then diluted to 4nM according to NextSeq manufacturers instructions. 1.5pM was loaded onto the Flow Cell with 1% PhiX library control. Libraries were sequenced by Illumina NextSeq 550 platform with single-end reads of 75 cycles according to the manufacturers instructions.