Reagents and workflow are optimized for efficient RNA isolation and recovery from Cell-Mate3D™ cultures. Protocol is derived from methods extracting RNA from polysaccharide rich tissues. 1,2,3
Cell-Mate3D™ has unique chemical and physical properties such that common RNA isolation kits are not suitable for RNA extraction.
Nanodrop spectra of isolated RNA from 7 individual Cell-Mate3D™ samples with Varying cell densities. The total RNA yield represented range from 1.8ug to 16.8ug.
Agilent Bioanalyzer electropherogram depicting high quality total RNA isolated from Cell-Mate3D™ culture followed by Qiagen® RNA clean-up. Note clearly delineated 28S and 18S peaks and minimal low molecular weight noise. rRNA Ratio 28S/18S=2.2
q-RT-PCR amplification plot demonstrating GAPDH expression. RNA was isolated from Cell-Mate3D™ Cultures and converted to cDNA. 200ng of cDNA per well was used in each q-RT-PCR reaction. Technical triplicates were performed. Average CT value was 15.5.