Xiaolian Gao
University of Houston, USA
Title: Advanced work flow for efficient multiplexing synthesis of genes in high fidelity – Next generation of gene synthesis
Biography
Biography: Xiaolian Gao
Abstract
This presentation will describe our synthetic biology project aiming a streamlining process of multiplex high fidelity gene synthesis using microchip oligo building blocks. This process features miniaturization, computation bioinformatics design, optimized work flow, low material consumption, long and high sequence accuracy, low error DNA constructs through efficient production process. Specifically, our work established a simple and easy to use flow column method (immobilized cellulose-binding-mutS column) to remove error-containing sequences from the final oligo gene-building blocks which are designed as such that they can be processed by ligation and PCR to give defined long (kb) DNA constructs. The reported workflow required about an hour of bench time for oligo processing, and attained less than 1 error per kb DNA, which is translated to ~80% success rate of full length EGFP (720 bp) gene cloning. The workflow hands more than ten genes in parallel. Has the potential for application in pathway gene cluster synthesis.