◊ Ensure sample homogeneity for accurate nucleic acid quantitation when using the NanoDrop ND-1000 spectrophotometer.
◊ Protect fluorescently tagged nucleotide conjugates from long exposures to light.
1. Laser Cut and Laser Capture Microdissection:
Isolate specific cells of interest using laser capture microdissection instrumentation from Arcturus according to the manufacturers protocol.
2. RNA Extraction and Isolation:
Perform RNA extractions and isolations using the Arcturus PicoPure RNA Isolation Kit or the Paradise Reagent System according to the manufacturers protocol. Expected total RNA yield will vary depending on starting material. Note: The quantity of total RNA extracted is dependent on several factors, including cell type and tissue quality. Certain cell types, such as monocytes, may yield extremely low RNA quantities, and may fall below the guidelines mentioned here. Please contact Arcturus Technical Support for questions related to RNA yield.
3. RNA Quantity and Purity Assessment:
This step is optional and may be performed if the starting cell number is ≥ 1000 cells. Total RNA can be quantitated, and the purity assessed, using the NanoDrop ND-1000 Spectrophotometer (figure 1a), while the quality of the total RNA can be determined using the RNA 6000 PicoLabChip on the Agilent 2100 bioanalyzer (figure 1b). RNA concentrations < 2ng/L or 260/280 ratios < 1.6, as determined by NanoDrop ND-1000, may indicate incomplete extraction, inefficient isolation, or co