Thank you for posting this protocol. This is very nice for us...

MS
Maria Solano
Sep 11, 2013
protocol Protocol: Single-cell Gene Expression Profiling of Mouse Stem Cells With Fluidigm BiomarkTM Dynamic Array
Thank you for posting this protocol. This is very nice for us out there trying to set the assay up.

How many technical repeats did you do per Taqman Assay primer pair?

How many technical repeats did you do per Biological replicate (ie cell)?



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Ana Sevilla Answered Sep 15, 2013

Department of Developmental and Regenerative Biology, Mount Sinai School of Medicine, USA

Hi Maria.
In the 96 Fluidigm Chip you have the option to interrogate 96 Taqman Assay Primers.
For single cell assays I would recommend you to do more biological replicates rather than technical replicates. I have run twice the same cDNA in two different Fluidigm Chips and the results were the same. The technique is pretty robust.
In our paper we interrogated 96 cells (biological replicates) per condition. For single cell analysis the more biological replicates you can afford the more robust statistic results you will get.
The size of the Taqman Assay Primers you will have to purchase will depend on the number of Chips you would like to run.
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