User:Paulina Alatriste/Notebook/UNAM Genomics Mexico 2011/2011/08/09
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Planning of the assembly of parts/Oligo design
-The parts come in six plasmids, we need to obtain all the fragments with PCR using oligos that have an overlapping region with other parts in order to perform Gibson assembly. -In a first step of ligation the parts that where synthesized separated into halves are going to be assembled. This is the case of PFOR and HydEF where each halve of the gene comes in different plasmids. -In a second step of ligation PFOR-HydA and HydEF-HydG are going to be assembled. -We have to design and order the oligonucleotides immediately to amplify the regions out of the plasmids. It is necessary to do this design consciously, because they need to have determined overlapping regions. -We need to send to the registry all the parts separated, in total 4 parts. -We need to find out what is the best: clean the PCR product and then isolate it from a gel band, or only isolate the product from a gel band.