TRIzol Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Protocol tips
- For low 260/230 readings the best approach is to try washing sample (precipitation) with ethanol to desalt it.

Publication protocol

Approximately 5 × 106 Sf9 cells were collected, and the total RNA was isolated following the manufacturer’s instructions using TRIzol reagent (Invitrogen, USA). RNA degradation and contamination were assessed using 1% agarose gels. RNA purity was detected using the NanoPhotometer® spectrophotometer (IMPLEN, CA, USA). An RNA Nano 6000 Assay Kit was used to assess the RNA integrity using the Agilent Bioanalyzer 2100 system (Agilent Technologies, USA). The total RNA of three biological replicates was mixed together, and 3 μg of mixed RNA was used for the RNA sample preparation. Sequencing libraries were generated using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (NEB, USA) following the manufacturer’s instructions.

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Manufacturer protocol

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