PicoPure™ RNA Isolation Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Can be used when there is a very less amount of starting material.

Upstream tips
- Can be used when there is a very less amount of starting material.

Publication protocol

RNA was isolated with the TRIzol®
reagent (Invitrogen) or with the ARCTURUS® PicoPure® RNA Isolation Kit (Applied Biosystems, Foster City, CA, USA) according to the manufacturer’s instructions. DNase digestion, cDNA synthesis, and qRT-PCR were performed as described previously15 using TaqMan primer probes (Applied Biosystems): glyceraldehyde 3-phosphate dehydrogenase
(GAPDH) (Hs99999905_m1), CXCR4 (Hs00237052_
m1), CXCR7 (Hs00664172_s1), FN1 (Hs00277509_
m1), GFAP (Hs00157674_m1), Iba-1 (Hs00610419_g1),
SNAI1 (Hs00195591_m1), SNAI2 (Hs00950344_m1),
TWIST1 (Hs01675818_s1), VIM (Hs00185584_m1),
CD133 (PROM1) (Hs00195682_m1), NES (Hs00707120_
s1), SOX2 (Hs00602736_s1), MSI1 (Hs00159291_m1),
ITGA5 (Hs00233743_m1), ITGB4 (Hs00236216_m1), and
CD3 (Hs01062241_m1). Fluorescence data were measured and converted into cycle threshold (Ct) data, and DCt values of each sample were calculated as Ctgene of interest− CtGAPDH. Relative gene expression was calculated with y=2(normalized CT nonmigrated−normalized CT migrated) = n-fold expression compared to slow-migrating cells/control. A DCt value of 3.33 corresponds to a one-magnitude-lower gene expression compared to GAPDH.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for PicoPure™ RNA Isolation Kit below.

Download manufacturer protocol

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