OT-2 Counter-Selection v2

This protocol is meant to select samples from one or more source plate(s) that has 2 different selecting conditions: one condition gives higher values and the other lower values (i.e different fluorescence values or OD600nm values to indicate growth/no growth). From these selected samples this protocol creates final plates with given reactive(s) (i.e media with different antibiotics or inductors) and inoculates samples from a source plate.In our laboratory this protocol has been used as part of the "High-throughput workflow for the genotypic characterization of transposon insertion library variants" also available in protocols.io to select variants to keep as glycerol stock and perform subsequent PCRs to assign a genotype. In particular, this protocol is used for selecting bacterial variants grown in two 96-well plates with different antibiotics by measuring OD600nm after overnight growth. Bacterial variants are selected based on growth/no growth and inoculated in final plates. Final plates consist of 96-well format plates with molecular biology grade water to perform subsequent PCR on selected samples and 2 stock plates containing 30% glycerol to perform bacterial stocks of selected samples. To run this protocol a python script for an Opentrons OT-2 robot and an excel file with several variables are needed . This protocol is a set of instructions or description of the LAP repository entry LAP-ColonyCounterSelection-OT2-2.0.0 You can find the script and complementary information for this specific version of the protocol in this LAP entry link and GitHub Link to LAP entry documents The major changes from previous version are: Change of name variableAPI Name Rack 15mL Falcon Reactiveswhich is called nowAPI Name Rack Falcon Reactives The program now accepts falcon tubes and tube racks of 15ml or 50ml. It does not accept mix tube racks. Description of robot and protocol setup in a separate protocols.io entry (Setting and Customizing OT-2 for LAP Entries) Sheets that corresponds to the map of values to compare now need to have the name of the rows and columns of the source labware