GSM5035224: 02_Mtb_pH5_24h; Mycobacterium tuberculosis; RNA-Seq
In three biological replicates, Mtuberculosis cultured in 7H9 rich media with 0.05% tyloxapol were washed 3x in sterile 1x PBS and diluted in 7H9 rich media with 0.05% tyloxapol buffered to either pH7 or pH 5.6 to an OD of 0.05 and grown in roller bottles in a 37 degrees C incubator for either 0 or 24 h.  After harvesting at 0 or 24 h, cells pellets were spun down and immediately frozen in liquid nitrogen and stored at -80 degrees C until RNA isolation . Cell pellets were transferred to a tube containing Lysing Matrix B, and vigorously shaken at max speed for 30 s in a FastPrep 120 homogenizer three times. This mixture was centrifuged at max speed for 1 min and the supernatant was transferred to a fresh tube. RNA was isolated using phenol-chloroform extraction method, followed by DNase treatment. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM5035224