OT-2 Modular Cloning Construct Assembly v2

This protocol is meant to create modular cloning constructs from different parts into a final plate and, optionally, perform the temperature profile needed to the assembly of the constructs. The output of running this script will be the final plate(s) with the constructs and the mix needed to perform that assembly, and the corresponding map(s) with the names of the constructs in their corrresponding well which will be given by the user in the input file. This protocol is run by using a LAP format script and its corresponding .xlsx file were different customizable variables such as the number of final combinations, volumes of transfer, type of plates, etc... Specifically, this protocol provides a set of instructions or description of the LAP repository entry LAP-MoCloAssembly-OT2-2.0.0. You can find the script and complementary information for this specific version of the protocol in the LAP entry link and GitHub link to LAP entry documents In our laboratory, this protocol has been used to perform plasmids using the Golden Standard modular cloning of levels 1 and level 2 The current version incorporates the following modifications: Description of robot and protocol setup in a separate protocols.io entry (Setting and Customizing OT-2 for LAP Entries) New variables added:Position Distribute Water, Touch Tip After Distributing Water, Change Tip in Water Distribution, Position Distribute Reaction Mix, Touch Tip After Distributing Reaction Mix, Change Tip in Mix Distribution, Position Distribute Acceptor/Module, Touch Tip After Distributing Acceptor/Module, Change Tip in Acceptor/Module Distribution, Max Volume Per Mix Tube In Shaker Iteration through all the DNA parts, first distributing acceptors and then modules, to all the final combinations that include those parts. Tips are being replaced every time volume is aspirated from the second reagent onwards when transferred to the mix tubes, avoiding contamination of the original reagents.