Category: Automation and High-Throughput Technologies

1346-B - Miniaturization of a Five Piece PIK3CA Plasmid Assembly

Monday, February 5, 2018
5:00 PM - 6:00 PM

The assembly of component DNA parts into a sequence-verified plasmid is a fundamental workflow within synthetic biology. Quickly moving projects through the iterative design, build, test, and learn phases is crucial in the ever-evolving market. As the scale and complexity of those projects increase, the DNA build phase can present hurdles in terms of cycle time and reagent cost. In this experiment, the Labcyte Echo® 525 Liquid Handler is used in combination with the NEBuilder HiFi® Assembly kit to accomplish the overall goal of accurately, quickly, and inexpensively constructing a circular plasmid from linear input pieces. In the experiment, we demonstrate the ability to reduce the NEBuilder assembly reagent volume on a complex five-piece assembly by forty-fold from the original recommended volume. In addition, the use of Echo-enabled colony qPCR for quality control and acoustic E. coli spotting for transformant plating are shown to be beneficial for the DNA assembly pipeline. Using the Echo 525 Liquid Handler, the cost and time required for DNA assembly can be dramatically reduced. 

Jared Bailey

Applications Scientist
Labcyte, Inc.
San Jose, CA

Jared's focus is on synthetic biology and DNA construction. He obtained his Bachelor’s degree from Otterbein University and his Master's degree from the University of Kentucky. He previously has worked at Allylix and Intrexon on the biosynthetic production of terpenes and isobutanol. Currently, Jared is working on the integration of the Labcyte Echo into synthetic biology workflows.