FastRNA™ SPIN Kit for Yeast

Protocol tips

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	Briefly, cell disruption was performed with 3×2 minute bursts, using a BioSpec Products Mini–Beadbeater–96 (Bartlesville, OK, USA) and purified RNA samples were flash frozen in liquid nitrogen before storage at –80°C.

Protocol tips
Briefly, cell disruption was performed with 3×2 minute bursts, using a BioSpec Products Mini–Beadbeater–96 (Bartlesville, OK, USA) and purified RNA samples were flash frozen in liquid nitrogen before storage at –80°C.

Publication protocol

otal RNA samples were prepared in duplicate from 8 samples using a FastRNA™ Yeast SPIN kit (MP Biomedicals, Santa Ana, CA, USA). Briefly, cell disruption was performed with 3×2 minute bursts, using a BioSpec Products Mini–Beadbeater–96 (Bartlesville, OK, USA) and purified RNA samples were flash frozen in liquid nitrogen before storage at –80°C. cDNA libraries were constructed using Illumina TruSeq stranded mRNA library preparation kits and sequenced on an Illumina HiSeq 2500 platform. TruSeq libraries of two additional P. pastoris strains expressing the β-carotene biosynthetic pathway, either regulated by the constitutive GAP promoter or the inducible AOX1 promoter, were also analyzed by RNA-seq; these samples were similarly drawn after 48 h growth on glucose and after 24 h of methanol induction, respectively. Additional libraries were prepared from BG10 strains expressing a variety of heterologous proteins, both intracellular and secreted. In total, 57 RNA-seq libraries were created and sequenced.

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Manufacturer protocol

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