Park Seongjin, Bujnowska Magda, McLean Eric L, Fei Jingyi
Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL, USA.
The College of The University of Chicago, Chicago, IL, USA.
Methods Mol Biol. 2018;1737:199-212. doi: 10.1007/978-1-4939-7634-8_12.
We present a method for the quantification of small regulatory RNAs (sRNAs) in bacteria, by combining single-molecule fluorescence in situ hybridization (smFISH), super-resolved single-fluorophore microscopy, and clustering analysis. Compared to smFISH imaging with diffraction-limited fluorescence microscopy, our method provides better quantification for short and abundant RNA (such as sRNAs) in a small volume of bacterial cells. Our method can also be directly used for the quantification of messenger RNAs (mRNAs).
我们提出了一种通过结合单分子荧光原位杂交（smFISH）、超分辨单荧光团显微镜和聚类分析来定量细菌中小调节RNA（sRNA）的方法。与使用衍射极限荧光显微镜的smFISH成像相比，我们的方法能更好地定量少量细菌细胞中的短且丰富的RNA（如sRNA）。我们的方法也可直接用于信使RNA（mRNA）的定量。
