RNAzol® RT

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- To reduce RNA degradation do not wash cells before the addition of RNAzol® RT. - To reduce DNA contamination, use sufficient amount of RNAzol® RT based on the area of the culture dish and not on cell number.	To reduce DNA contamination extend the incubation time to 15 minutes before sedimentation.

Upstream tips
- To reduce RNA degradation do not wash cells before the addition of RNAzol® RT. - To reduce DNA contamination, use sufficient amount of RNAzol® RT based on the area of the culture dish and not on cell number.
Protocol tips
To reduce DNA contamination extend the incubation time to 15 minutes before sedimentation.

Publication protocol

RNAzol RT (Molecular Research Center,
USA) was used to extract total RNA from glioma
tissue, adjacent normal tissue, glioma cell lines,
HBMECs and exosomal sources24. qRT-PCR for
human-specific repeat sequences was performed. To identify the gene expression of HBMECs after treatment with
A172-Exo and shA172-Exo, 8 × 105
HBMECs were seeded onto six-well plates and cultured with fresh medium containing 100 μg/mL exosomes for 48 h. The primers were obtained from Gene-Pharma (Shanghai, China), and the following
human primers were used: linc-POU3F3, basic
fibroblast growth factor (bFGF), bFGF receptor
(bFGFR), vascular endothelial growth factor A
(VEGFA), VEGF receptor 2 (KDR), transforming growth factor beta 1 (TGFB1), intercellular
adhesion molecule-1 (ICAM-1), and angiogenin
(Angio). GAPDH was used to normalize lincRNA in the tissue, cells and exosomes. The primer
sequences used in this study are summarized in
Table II. Each sample was analyzed in triplicate
for yield validation. The 2−ΔΔCt method was used
to determine the relative quantification of gene
expression levels.

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Manufacturer protocol

Download the product protocol from Molecular Research Center, Inc. for RNAzol® RT below.

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