Developing Custom Picking Software Using the Genetix SDK Presenting Author: Steven E. Wilson U.S. DOE Joint Genome Institute2800 Mitchell Drive, Bldg. 100 Walnut Creek, CA 94598 sewilson@lbl.gov(925) 296-5769 Automated colony picking has allowed DOE s Joint Genome Institute to inoculate upwards of 215 384-well plates per day. JGI currently attains a ratio of about 2.5 destination plates per bioassay tray (~1000 colonies per bioassay tray) and 2 or fewer failed wells per plate. The picking setup screen can also be overwhelming for new users that do not know how to navigate the user interface.The Genetix software developer s toolkit (SDK) allows programmers to create imaging and picking routines that suite the specific needs of their clients. The JGI has taken advantage of the SDK by creating a program that shows the machine operators only what they need to see on a plate to plate basis, uses a special calibration routine and implementing a background subtraction routine that increases the contrast between the agar media and the colonies. Used together, the JGI will stabilize the number of failed wells and increase the number of colonies that are imaged per bioassay tray. With the implementation of the custom software, JGI hopes to reduce reagent and plastics costs, increase picking efficiency and streamline the picking process.

I. AN OVERVIEW OF THE LIBRARY SUPPORT PROCESS USED AT THE JOINT GENOME INSTITUTE PRODUCTION GENOMICS FACILITY Cathe Adam, Karli Ikeda, Sandy Huynh, Jayadevi Krishnakumar, Jim Fey, Diane Bauer, Sanna Anwar, Mansi Chovatia, Nicole Shapiro, Miranda Harmon-SmithLibrary support is the first step in production sequencing at the Joint Genome Institute s (JGI) Production Genomics Facility (PGF). The goal of library support is to generate 384 well glycerol stock plates of each library for a given project. These plates are then passed on to the Sequencing Prep group for Rolling Circle Amplification. Transformation stocks of the 3, 8, and 40kb libraries for a given project are received into library support from cloning technology. The stocks are stored at -80oC until plated. Libraries are plated according to priority. The colonies produced from a plating event are then picked using robotic colony pickers and are inoculated into 384-well destination plates containing LB/glycerol + antibiotic. The destination plates are grown overnight and visualized on a Spectramax Optical Density reader to identify wells without growth. Wells containing no growth are aspirated and replaced with colony growth from a "fix source" plate generated from the same library. The destination plates are then sealed and stored at -80oC until sent to the Sequencing Prep area for Rolling Circle Amplification (RCA). This poster will elaborate on these steps in the library support process and detail further the project tracking and management, plating and picking procedures and the instruments that are utilized throughout these steps.

Library support is the first step in production sequencing at the Joint Genome Institute's (JGI) Production Genomics Facility (PGF). The goal of library support is to generate 384 well glycerol stock plates of each library for a given project. These plates are then passed on to the Sequencing Prep group for Rolling Circle Amplification. Transformation stocks of the 3, 8, and 40kb libraries for a given project are received into library support from cloning technology. The stocks are stored at -80oC until plated. Libraries are plated according to priority. The colonies produced from a plating event are then picked using robotic colony pickers and are inoculated into 384-well destination plates containing LB/glycerol + antibiotic. The destination plates are grown overnight and visualized on a Spectramax Optical Density reader to identify wells without growth. Wells containing no growth are aspirated and replaced with colony growth from a "fix source" plate generated from the same library. The destination plates are then sealed and stored at -80oC until sent to the Sequencing Prep area for Rolling Circle Amplification (RCA). This poster will elaborate on these steps in the library support process and detail further the project tracking and management, plating and picking procedures and the instruments that are utilized throughout these steps.