• EpCAM binding docks are cloned and confirmed.
The gene sequences of EpCAM binding docks are synthesized particularly and separately. The confirmation electrophoresis and PCR afterwards were found correct. BBa_K772001 and BBa_K772002 are fixed together via 3A assembly method. After transformation into E.coli, the gel bands match with their original length; therefore we proceeded to the next step: Sequencing.
• Binding docks are sequenced and found correct. Sequence results overlap with the original sequences and show that these two parts are assembled correctly and ready to test for the synthesis and its function.
• Binding docks are synthesized and are able to bind to the EpCAM.
After the correct ligation and cloning procedures, we extracted both bacterial and tumor cell protein ingredients via performing sonication. Affinity Gel Chromatography is performed on bacterial protein extract in order to isolate our EpCAM binding docks. These docks were exposed to tumor cell extract; thus the binding would occur. SDS-PAGE is done aftermath on the last solution.
Applications of BBa_K772002