GBM23 - MetabolicEngineeringGroupCBMA/MetabolicEngineeringGroupCBMA.github.io GitHub Wiki
This document contain notes and links pertaining to the master course Genética e Biotecnologia Molecular 2023/2024 given to students attending Mestrado em Bioquímica Aplicada (MBQA) or Mestrado em Genética Molecular (MGM).
You need to bring a laptop computer to this class, and you also need to have a Google account and the ApE plasmid editor app.
Your task is to assemble the sequence of the ATF1 expression vector pTA1_TDH3_ScATF1_PGI1 using the Yeast Pathway Kit (YPK). The YPK is an assembly protocol developed by the MEC group (publication).
Assembly of the pYPKa_A_ATF1 vector
Your first task is to assemble the pYPKa_A_ATF1 vector. Follow these instructions to do this.
When you have a candidate for the pYPKa_A_ATF1 vector sequence, calculate the cSEGUID checksum for the raw sequence using SEGUID calculator. Modern versions of ApE also have this functionality.
When you are done, fill in your results in the Google spreadsheet. The link can be found in Blackboard
Assembly of the pTA1_TDH3_ScATF1_PGI1 vector
Your second task is to assemble the pTA1_TDH3_ScATF1_PGI1 vector. You need the pYPKa_A_ATF1 plasmid sequence that you assembled in the previous step to do this. T This is more complex as it is made by homologous recombination of four DNA fragments.
Follow these instructions.
When you are done, fill in your results in the Google spreadsheet.
Yeast transformation
Yeast transformation Protocol.