The Microbial Transformation tool allows the user to select one or more plates of DNA materials and to select a microbial material for transformation. The tool generates a worklist guiding the addition of DNA materials to a new plate of the selected microbial material.
Input: Plates containing DNA materials or a transformation reaction map.
Output: A destination plate containing the microbial material, a transformation reaction map, and a worklist guiding the transfer of volumes from the plate(s) of DNA materials to the destination plate of the selected microbial material.
You can find more detailed definitions of the terms mentioned in this article in the glossary.
(required) The plate(s) must already exist in inventory (Inventory > Plates)
If the plate does not exist yet, use the header to go to Inventory > Plates > ‘New Plate’ or ‘Upload’ to create a new plate
(required) The plate(s) must be hydrated
If the plate is not hydrated, go to Inventory > Plates > double-click on the plate to go into the plate record view > ‘Update’ > ‘Update Plate’ > set ‘Plate Hydration’ to ‘Rehydrate’ > set the ‘Target Volume’ > Submit
(required) The microbial material must exist in the microbial materials library (Materials > Microbial Materials)
If the material does not exist yet, use the header to go to Materials > Microbial Materials > ‘New Material’
(optional) The user can create a transformation reaction map via the Create Reaction Map tool
Create Reaction Map tool under Tools > Tool Library
‘Inputs’ = the microbial material, the plasmid
‘Output’ = the microbial material containing the plasmid
‘Reaction Map Type’ = ‘Clonal_Transformation’
(optional) The user can normalize the concentration of the DNA materials in one or more plates to the same target concentration via the Normalization Planning tool and Execute Worklist tool
Normalization Planning tool and Execute Worklist tool under Tools > Tool Library
You can find the Microbial Transformation tool under Tools > Tool Library. Click ‘Launch Tool’ to start using the tool.
The tool will walk you through several steps to generate a worklist for microbial transformation. You need to complete each step before proceeding to the next. After completing a step, the completed step mark (circle) will turn green with a check mark. You can always use the ‘previous’ button to go back to any previous step.
1. Selecting Plates: In this step, you need to select the plates containing the DNA material that you wish to transform into a microbial material.
1a. Select Transformation Reaction Map: This step is optional. If a transformation reaction map is selected, the microbial material will be auto-selected in the next step.
1b. Select Plates From Inventory: This step is required. Click on ‘Select Plates’ button to select one or more plates (from the inventory) containing DNA materials that you wish to do the transformation.
After selecting the desired plate(s), you can view the table of selected plates. To change the selections, click ‘Change Plates’, or click on the red trash can icon above the table to remove all selected plates from the list. To remove a single plate from the list, click on the red trash can icon at the right of that plate’s row. Click on the eye icon at the left of a plate’s row in order to see the plate map and listed contents. When satisfied with the selections, click ‘Next’.
2. Transformation Prep: In this step, you need to select a microbial material, specify the amount of DNA material and microbial material for the transformation, and optionally specify the name and the type of the destination plate that stores the transformation reaction.
2a. Microbial Material Information: If you selected a transformation reaction map in the previous step, a microbial material will be auto-selected for you in this step. Otherwise, click ‘Select Material’ and select a microbial material from the inventory. You can use the search function within the pop-up window to search for a specific material.
2b. Transfer Volume: After selecting a microbial material, you need to specify the amount of DNA material and microbial material needed for the transformation reaction.
2c. Destination Plate Information: Enter a plate name and select a plate type. The default plate name and type are ‘Destination Plate 1’ and ‘Generic 96 Well Plate’ respectively. We recommend selecting a descriptive name for your destination plate. Click ‘Next’ after you are done with the ‘Transformation Prep’ step to go the next step.
This step allows you to review the transformation worklist, which contains liquid handling instructions for transferring volumes from the plate(s) of DNA materials to the destination plate. Rename the worklist and click ‘Generate Worklist’.
You will then be notified that the microbial transformation is completed and there are links to the new microbial plate(s) that stores the transformed cells, a transformation worklist, and a transformation reaction map which will be used to validate the transformation reaction when the transformation worklist is executed (via 'Execute Worklist' tool).
If you click on the link to the microbial plate, you will see that the plate only contains the volumes of the selected microbial material.
The transformation reaction map displays the inputs of microbial material and plasmids as well as the output of microbial material containing the plasmids
The transformation worklist contains instructions for transferring volumes from the plate(s) of DNA materials to the destination plate that should already contain the microbial material. Click on ‘Export Worklist’ to export the worklist in the preferred format of different liquid handlers. After executing the worklist in the lab, use the Execute Worklist tool under Tools > Tool Library to update the plate in the inventory.