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Bacterial Conjugation Tool
Bacterial Conjugation Tool

Transfer DNA between two cells

Jota Hurtado avatar
Written by Jota Hurtado
Updated over a week ago

This tool allows users to transfer DNA materials between two cells.

Input: Plates or tubes of donor cells (microbial materials containing plasmids) and recipient cells (microbial materials containing none or some plasmids). Note that materials in the input containers of the donor cells will be consumed entirely.

Output: Plates or tubes of recipient cells with plasmids coming from donor cells.

You can find more detailed definitions of the terms mentioned in this article in the glossary.

Prerequisites:

  • (required) The donor plate(s) or tube(s) must already exist in inventory (Inventory > Plates/Tubes).

  • If the plate (or tube) does not exist yet, go to Inventory > Plates > ‘New Plate’ or ‘Upload’ to create a new plate. Inventory > Plates > ‘New Tube’ or ‘Upload’ to create a new tube.

You can find the ‘Bacterial Conjugation’ 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 bacterial conjugation. 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 checkmark. You can always use the ‘previous’ button to go back to any previous step.

1. Select Donor Cell: Click ‘Select Plates’ or ‘Select Tubes’ to select one or more plates or tubes from inventory dialog by checking the box(es) next to the desired plate(s) or tube(s). Click ‘Select Plates’ or ‘Select Tubes’ to confirm your selection.

After selecting the desired plate(s) or tube(s), you can view the table of selected plates or tubes. 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’ to go to the next step.

2. Select Recipient Cells: Click ‘Select Material’ to choose one or more ‘Microbial Material’ from inventory to add to the selected material by checking the box next to the desired material. Click ‘Select Material’ to confirm your selection.

After selecting the desired recipient material, you can view its information. To change the selection, click ‘Change Material’, or click on the red trash can icon next to the ‘Change Material’ button to remove the material.

You need to specify a transfer volume of recipient cells to transfer to the selected donor cell's container. Then you need to enter a name and select the type for an intermediate container, which will hold the microbial material prior to transferring. Optionally you can manually generate a barcode for the intermediate container or you can just leave the ‘Generate Barcode’ checkbox selected so the barcode is generated automatically. Finally, you need to specify the dead volume expected for your liquid handler and selected container type.

3. Review Worklist: Review your generated worklist to be sure it embodies the transfers you expect.

4. Submit: Submit your work and receive the following message for successfully tracking bacterial conjugation. Note that you can go to both the worklist or the reaction map to inspect them if you wish.

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