3 articles published in JoVE
Characterize Disease-related Mutants of RAF Family Kinases by Using a Set of Practical and Feasible Methods Jiajun Yap1, Jimin Yuan1, Zi Heng Tee1, Xuchao Huang1, Wan Hwa Ng1, Jiancheng Hu1,2 1The Laboratory of Cancer Signaling, Division of Cellular and Molecular Research, National Cancer Centre Singapore, 2The Cancer and Stem Cell Program, Duke-NUS Medical School In this article, we presented a set of practical and feasible methods for characterizing disease-related mutants of RAF family kinases, which include in vitro kinase assay, RAF co-activation assay, and complementary split luciferase assay.
Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format Muhammad Khairul Ramlee1, Jing Wang1, Alice M. S. Cheung1, Shang Li1 1Cancer & Stem Cell Biology Programme, Duke-NUS Medical School The genotyping technique described here, which couples fluorescent polymerase chain reaction (PCR) to capillary gel electrophoresis, allows for high-throughput genotyping of nuclease-mediated knockout clones. It circumvents limitations faced by other genotyping techniques and is more cost effective than sequencing methods.
Detection of RNA-binding Proteins by In Vitro RNA Pull-down in Adipocyte Culture Qianfan Bai*1, Zhiqiang Bai*1, Shaohai Xu3, Lei Sun1,2 1Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, 2Institute of Molecular and Cell Biology, Singapore, 3Division of Bioengineering, School of Chemical & Biomedical Engineering, Nanyang Technological University An RNA pull-down protocol is optimized here for detection of interactions between RNA-binding proteins (RBPs) and noncoding as well as coding RNAs. An RNA fragment from androgen receptor (AR) was used as an example to demonstrate how to retrieve its RBP from lystate of primary brown adipocytes.