MOESM2 of A rapid preparation procedure for laser microdissection-mediated harvest of plant tissues for gene expression analysis

Additional file 2. RNA and qPCR data. RNA quantity, quality and duration of storage in a desiccator at 4 °C prior to LM. Gene expression data including Cq values, calculations and p-values. Gene-specific primer sequences, expected amplicon sizes and qPCR amplification efficiencies calculated based on standard curves of diluted cDNA. Size separation of qPCR amplicons on a 2% agarose gel under constant voltage set to 100 (nucleic acids were stained with GelRed and visualized using the ChemiDoc MP Imaging System (Bio-Rad). The GeneRuler 50 bp Ladder (Life Technologies) was included for size estimation). Melt peaks of qPCR amplicons generated by plotting the negative derivative of the change in fluorescence intensity as a function of temperature.

补充文件2：RNA与定量聚合酶链反应（qPCR）数据集。本数据集包含RNA的浓度、质量，以及在4℃干燥器中储存至激光显微切割（LM）前的时长；基因表达数据涵盖定量循环阈值（Cq）值、计算结果与显著性p值；基因特异性引物序列、预期扩增子长度，以及基于稀释cDNA标准曲线计算得到的qPCR扩增效率。qPCR扩增子的尺寸分离实验：以恒压100V在2%琼脂糖凝胶中进行电泳分离，其中核酸采用GelRed染色，通过ChemiDoc MP成像系统（伯乐Bio-Rad）完成可视化，实验中加入GeneRuler 50 bp DNA分子量标准（Life Technologies）用于分子量估算。qPCR扩增子的熔解峰则通过绘制荧光强度变化的负导数随温度变化的曲线获得。