[BME Colloquium] High-throughput DNA synthesis for synthetic biology applications
  • Duhee Bang, Ph.D. (Yonsei University)
  • SLS Colloquia / Oct. 16th 04:00 pm / EB1 E104
Abstract
 

Screening highly complex sequence gene libraries are powerful tool for fine mapping of sequence-function relationship. Typically, these libraries are made by error-prone PCR, random shuffling, or assembly of synthetic oligonucleotide variant libraries. By careful scrutinization of individual variant library, we can possibly delineate the crucial role for each sequence variation, which leads to change the activities of protein. However, as the number of the library sequences exceeds hundred clones, sequence verification of individual variants is subjected to tedious procedures, and become limited in scalability. As a result, only a few clones after the selection have been subjected to further analysis via conventional cloning and Sanger sequencing. Although the characterization of a few ‘selected’ clones has shown great utility for the discovery of sequence information with top functional characteristics, small number of clones would not represent full-spectrum of sequence space that contains cryptic mutations that are potentially adaptive at a given environmental stress. In the presentation, I present a method that enables profiling of full-spectrum of gene variant sequences.