Part:BBa_K1456003:Design

Superoxide dismutase-1 (SOD-1)

We amplified the SOD1 genes using CMV forward and SV40 reverse primers and achieved results by viewing band at 526bp.
cDNA obtained from HEP 2G cells have been used to perform PCR to synthesize our desired inserts. EcoRI and BamHI were the choice of enzymes to digest our inserts once their purification was accomplished using phenol chloroform method. Transformation using the DH5α strain was the next step after ligating our inserts with the pTRE vectors. Colony PCR procedure was conducted using the CMV forward and SV40 reverse primers to ensure that our inserts were cloned correctly. Below are the results of our Colony pcr procedure:

According to the forth colony, our results show that sod1-ptre vector has been inserted correctly.
In order to prove that our cloning has been carried out correctly we used the restriction-digest process and achieved the expected results. We cotransfected our vector with HEK 293 T cell line. After incubating our cells we performed western blotting and controlled the cell expression of our SOD1 genes.

Assembly Compatibility: