Week 3: Automation

Class Assignment

Python Script

Uploaded to form and shared with lab.

Post-lab questions

I investigated the paper “Slowpoke: An Automated Golden Gate Cloning Workflow for Opentrons OT-2 and Flex” as I was interested in using GGA to assemble my engineered phage satellite for my final project and was curious as to how I could integrate an Opentrons workflow into the process. The paper tested two GGA kits, the MoClo Yeast Toolkit (YTK) and the SubtiToolKit (STK), with GFP as the reporter for both, then transformed the assembled plasmids into E. coli and Saccharomyces cerevisiae (yeast) or Bacilius subtilius based on the according kit. The authors found high assembly and transformation efficiencies using the opentrons to complete the protocols (17/17 colonies and 8/13 colonies contained the GGA construct with GFP using the YTK and STK kits respectively).

This paper shows promise for using the opentrons to run the protocols of other GGA kits for my final project. I also plan to use opentrons for phage plating, particularly for spot tests covering the plate in a webbed lysis form that can be flooded to increase the titer of phage and phage satellites, and have started working on a Python script for such applications.