Preparation of Standards from Purified DNA Amplicons and Treatment of MCF7 p53-Venus Cells
1:53
Preparation of Lysis Buffer for Cell Sorting
4:15
Cell Sorter Setup, Cell Harvesting, and Sorting
5:47
Exonuclease Treatment and Sort Quality Control Using qPCR
7:02
Gene Expression Measurement Using Microfluidic qPCR
9:03
Results: Gene Expression Profiling in Single Cells Using FACS Analysis and qPCR
10:09
Conclusion
Transcript
The overall goal of this procedure is to quantify the levels of mRNA in individual cells using microfluidic qPCR with internal standards. This method can help answer key questions in the field of single cell biology, such as whether cell populatio
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We describe a method to sort single mammalian cells and to quantify the expression of up to 96 target genes of interest in each cell. This method includes the use of internal qPCR standards to enable the estimation of absolute transcript counts.