Single-Cell Real Time Analysis
This service includes advice and guidance in the generation of single cell preparations, execution of cell viability, cell diameter and cell buoyancy measurements [all essential for a successful single cell capture] and processing of a sample from a single cell suspension to the generation of archive cDNA isolates generated from single cells.
Users need to register their samples through iLab. Please go to the Customer Portal and make your request.
How to bring your samples:
Samples should be brought in a 96 well plate at a concentration range 5ng/ul to 50ng/ul in a total volume of 10ul. The 260/280 ratio for the RNA should be no less than 1.5.
THINGS TO CONSIDER
The range of total RNA that can be used in a 5-µL reverse transcription reaction
is 2.5 pg to 250 ng. However, the success with a given sample in qPCR will depend on the level of gene expression for the genes of interest, the percentage of mRNA in the total RNA, and the number of cycles of preamplification performed prior to qPCR.
In general, for total RNA input in the range of 2ng to 250ng, 10-14 cycles of preamplification should be sufficient. For a total RNA input less than 2 ng, increasing the number of preamplification cycles to 18-20 may improve performance.