|Jackson Jr, John|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 1/27/2009
Publication Date: 1/27/2009
Citation: Yoder, A., Pajak, L., Jackson Jr, J.S., Hughes, S.R. 2009. Creating libraries for commercial yeast strains through miniaturization of cloning and transformations using the BioRAPTR FRD microfluidic workstation [abstract]. LabAutomation. Poster #12. p. 49.
Technical Abstract: The ability to miniaturize molecular reactions can lead to significant cost savings when creating libraries of thousands of clones. For this application Beckman Coulter partnered with the USDA to provide a low-volume automated solution for library cloning for use in the development of yeast strains to convert biomass to ethanol and to express proteins or valuable coproducts, such as insecticides and pharmaceuticals. In this experiment, the Gateway LR Clonase II reaction was miniaturized from a full-scale reaction to 10% of the original volume using a full factorial design of experiments (DOE). The BioRAPTR FRD microfluidic workstation was used to dispense the vector, insert, and LR Clonase II reagents. The second part of the experiment tested the feasibility of miniaturizing the downstream transformation of chemically-competent bacteria. Results from miniaturization of the LR Clonase reaction showed no significant differences in the colony counts of the transformants. Similarly, there was no significant difference in the colony counts for the miniaturized transformations.