CRISPRi-seq of SPV_0476 complementation: no dCas9 induction
Genomic DNA was isolated from bacterial cultures grown in CY medium until OD=0.3 in presence or absence of aTc (dCas9 inducer). Cell were harvested, lysed using 0.025% Deoxycholate for 20min at 30C. DNA was precipitated with isopropanol and dissolved in TE buffer. Illumina adapters were brought by amplificating the sgRNA region by PCR using specific primers with adapters. Fragments were extracted on agarose gel for correct size selection and further prepared for injection into MiniSeq.