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High-throughput workflow for the genotypic characterization of transposon library variants v1
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This is a workflow for the genotypic characterization of transposon library variants. It has been developed using an open-source Opentrons OT-2 robot, BLASTN for genomic annotations and modular sub-protocols (e.g., PCR sample preparation, OT-2 volume transfer, OT-2 counter selection, etc) that can be used for other tasks, thus providing a general-purpose pipeline. All steps follow a 96-well plate format for high-throughput analysis. The protocol is described for the characterization of transposon library variants generated with SEVA-Sib pBAMD1-x and pBLAM1-x plasmid sets that follow Standard European Vector Architecture (SEVA, https://seva-plasmids.com) and can be amplified with the standard PS1-PS6 primers. After the description of the protocol we present the results of an example generated at our laboratory (https://biocomputationlab.com) using the soil bacterium Pseudomonas putida KT2440 as acceptor strain.
Title: High-throughput workflow for the genotypic characterization of transposon library variants v1
Description:
This is a workflow for the genotypic characterization of transposon library variants.
It has been developed using an open-source Opentrons OT-2 robot, BLASTN for genomic annotations and modular sub-protocols (e.
g.
, PCR sample preparation, OT-2 volume transfer, OT-2 counter selection, etc) that can be used for other tasks, thus providing a general-purpose pipeline.
All steps follow a 96-well plate format for high-throughput analysis.
The protocol is described for the characterization of transposon library variants generated with SEVA-Sib pBAMD1-x and pBLAM1-x plasmid sets that follow Standard European Vector Architecture (SEVA, https://seva-plasmids.
com) and can be amplified with the standard PS1-PS6 primers.
After the description of the protocol we present the results of an example generated at our laboratory (https://biocomputationlab.
com) using the soil bacterium Pseudomonas putida KT2440 as acceptor strain.
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