RNeasy Micro Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- This kit can be used for the isolation of RNA from up to 5x10^5 cells, or up to 5 mg of tissue.		- Always blank the spectrophotometer with your elution buffer. - When measuring 260 nm absorbance, a unit of 1 on this scale equals to 40 ng of RNA.

Upstream tips
- This kit can be used for the isolation of RNA from up to 5x10^5 cells, or up to 5 mg of tissue.
Downstream tips
- Always blank the spectrophotometer with your elution buffer. - When measuring 260 nm absorbance, a unit of 1 on this scale equals to 40 ng of RNA.

Publication protocol

RNA was isolated from cell lysates by the RNeasy Micro Kit (Qiagen), and cDNA was synthesized from 1 ug RNA, using SuperScript III (Invitrogen) and Random Primers (Invitrogen), as per the manufacturer's protocol. Quantitative real-time PCR (qPCR) was carried out on StepOne Real Time PCR system (Applied Biosystems) using gene-specific primers designed with Primer-BLAST (Supplementary Table S5B-C) and synthesized by IDT Technologies. Validation of RNA-seq results was carried out using TaqMan Universal PCR Master Mix II with uracil-N-glycosylase (Applied Biosystems) and Universal ProbeLibrary System probes (Roche) following manufacturer's protocol. Validation of siRNA-/shRNA-mediated knockdown was carried out using Fast SYBR Green Master Mix (Invitrogen), as per the manufacturer's protocol. qPCR data were analyzed using relative quantification method and plotted as average fold-change compared to the control. GAPDH was used as an internal reference. For qPCR validation studies, GraphPad Prism software was used to perform linear regression and calculate R2 correlation coefficients.

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Manufacturer protocol

Download the product protocol from Qiagen for RNeasy Micro Kit below.

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