MC3T3-E1 single cell qPCR results (Biomark, Fluidigm)

Fluidigm Biomark qPCR raw data was pre-processed using Fluidigm Real-Time PCR Analysis software. Failed assays were removed. The default quality threshold cutoff (0.65) was used to identify potential artifacts. The baseline correction was to linear, and the Ct threshold detection method was set to Auto (Detectors). The Fluidigm Singular Analysis Toolset v3.6.2 was used to remove outliers using the IdentifyOutliers() command.

Cell types used for this study:
Wildtype MC3T3-E1 at differentiation stages T0, T3, T10, T14 and T21, Satb2-mutant cell lines C8 at T0 and C9 and T0 and T14.

本研究采用Fluidigm实时定量PCR分析软件，对Fluidigm Biomark平台的实时定量聚合酶链反应（quantitative PCR, qPCR）原始数据进行预处理：首先移除失败的检测反应，以默认质量阈值截点（0.65）识别潜在的检测伪影；基线校正采用线性校正模式，Ct阈值检测方法设置为自动检测器模式。随后通过Fluidigm Singular Analysis Toolset v3.6.2软件的IdentifyOutliers()命令剔除异常值。本研究使用的细胞类型如下：分化阶段分别为T0、T3、T10、T14及T21的野生型MC3T3-E1细胞，以及Satb2突变细胞系C8（T0阶段）、C9（T0及T14阶段）。