BuG@S Cy5 RNA labelling - BuG@S Cy5 RNA labelling

Type
labeling
Description
2-10ug of RNA sample was placed in a microfuge tube (0.5mL) with 3ug of random primers (1uL) and made up to a final volume of 11uL with H2O (DNase and RNase free, molecular biology grade). The RNA was then heated to 95C for 5min, snap cooled on ice and centrifuged. 5uL First Strand Buffer (5x), 2.5uL DTT (100mM), 2.3ul dNTP's (5mM dA/G/TTP, 2mM dCTP), 1.7uL of Cy5(1mM) and 2.5uL of SuperScript II (200U/uL) were added to make a final volume of 25uL. The solution was incubated in the dark at 25C (room temperature) for 10min and then at 42C in the dark for 90min.
Parameters
RNA quantity, fluorescence label, labelled extract yield
Links
All experiments using protocol BuG@S Cy5 RNA labelling: (E-BUGS-112, E-BUGS-119)