miRCURY LNATM Universal RT microRNA PCR (Exiqon) v1 (protocols.io.utbewin) [dataset]

Malgorzata Guz
2018 protocols.io  
1 Performing an RNA dilution series can be used to determine that input amounts are within the linear range of the system, and to ensure a good level of signal, with no sign of inhibition. Increasing the RNA volume to increase signal levels can have the opposite effect and result in less signal due to carry-over of more inhibitory compounds into the RT-PCR reaction. Standard methods for measurement of the RNA yield and quality are inappropriate for use with biofluid samples. The presence of
more » ... ier RNA in these samples makes measuring the low levels of endogenous RNA by OD260 impossible. Even if carrier was not included during the isolation, the RNA concentration in the eluate would still be too low for reliable OD260 quantification on a NanoDrop or other spectrophotometers. Therefore, an alternative way of monitoring yield and normalizing sample input has to be used. Exiqon recommends using RNA amounts based on starting volume rather than RNA quantity. RNA i nput -opti mi za ti on.xl sx RNA i nput -opti mi za ti on.xl sx mi croRNA-se rum-pl a sma -gui de l i ne s.pdf mi croRNA-se rum-pl a sma -gui de l i ne s.pdf RNA input -optimization
doi:10.17504/protocols.io.utbewin fatcat:7szr44dwdjbkzgc2mik7es7rge