Meta-transcriptomic sequencing of four-species co-cultures
Exponential cultures of Novosphingomonas sp. G5, Pseudomonas sp. G7, Flavobacterium sp. G16 and Microbacterium sp. D12 were adjusted to an optical density at 600 nm of 0.15 in tryptic soy broth (TSB) medium. One milliliter of each culture and a mixture of them, which contains 250 ¦ÌL of each, were then inoculated into 50 mL fresh TSB medium. After 12 h incubation at 25 ¡æ with shaking (200 rpm), cells were collected by centrifugation at 4000¡Á g. Total RNA were extracted with TRIzol reagent (Invitrogen) following the manufacturer¡¯s instructions. RNA-seq libraries constructed with co-culture RNA and the control RNA, which is a mixture of one microgram of each single species RNA, were sequenced by HiSeq 2500 platform with standard protocols (BGI, China).
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