title: RNA-seq of P. pacificus vul-2 mutants
design: Culture populations were allowed to grow until their food was exhausted, immediately after which the cultures were processed for sequencing.  Five mixed-stage plates were washed with 40 ml M9, centrifuged immediately at 1300 g for 4 min,  rinsed with 40 ml 0.9% NaCl treated with 40 µl ampicillin and 40 µl chloramphenicol and shaken  gently for 2 hours, and finally concentrated into a pellet by centrifugation and immediately frozen  in liquid nitrogen. NEBNext Ultradirectional RNA Library Kit (Cat # E7420L) was used to prepare libraries.