Team:Valencia UPV/Project/modules/methodology/flowchart

Project > Modules > Methodology > Cloning > Flowchart

All the parts we used were either obtained by PCR or ordered by GBlocks. Sequences need have the appropiate flanking sequences so they can be ligated into a pUPD vector and transformed into E. coli DH5alpha. Transformed cells are cultured in solid LB plates supplied with ampicilin and grown at 37°C. Colonies are picked and grown in liquid LB with shaking at 37°C. Minipreps are done from liquid culture and digestions were performed to check whether they are correct. Parts are then sequenced to ensure no mutation occurred during the PCR process.

Parts can be assembled into transcriptional units in an alpha vector provided that they follow the GoldenBraid 2.0 grammar. Once transcriptional units are constructed an iterative loop of binary assemblies can be performed using alpha and omega vectors in order to obtain the final construction.