Advanced Synthesis of Barcoded Beads in High-throughput scRNA-seq
Zhen Li ’20, Nanjing, China
Majors: Biology; Cognitive Science
Due to the increasing needs of human genome study, research in high-throughput and low-cost scRNA-seq platform has been pushed further to the forefront. Instead of averaging the results of multiple cells via traditional cell sequencing, single-cell RNA sequencing could narrow down the result to single-cell mutation and provide us explicit details for further bioinformatics analysis, especially for cancer diagnosis. Despite the fact that previous research has mainly been done with the streptavidin-coated beads and hydrosol-based beads, magnetic beads coated with carboxyl groups were chosen for further synthesis due to their outstanding high-temperature resistance and PCR stability. Even though ligation-based barcoding synthesis has been the most popular synthesis method, it has reached its bottleneck in the past few years. In order to achieve a higher cDNA capture efficiency, this research tries to improve the ligation-based synthesis with extension-based synthesis methodology via three rounds of split-pool synthesis. Reverse complement oligo FAM coated with fluorescence was applied to the research product for further quality checks. Such methodology provides us a higher resolution of cellular differences and a better understanding of the function of an individual cell in the context of its microenvironment.