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The Cibacron blue biomimetic affinity chromatography will result in a high…

- - - - Lab 4- Determination of LDH-A Concentration via a Colorimetric LDH Assay
        
        Lab 5/6- Conclusive Determination of LDH-A Product and Determination of Enzyme Viability and Size via Western Immunoblotting
    - - Lab 7- Determination of Enzymatic Activity via a Spectrophotometer Analysis of NADH Production during an LDH-A reaction with lactate and NAD+
        
        Lab 7- Determination of the inhibition type exhibited by the LDH-Inhibitor on LDH-A (Research Question B only).
- - - - Lab 10- Purification and Analysis of the Amplified LDH-A Expression gene via Gel Electrophoresis. Preparation for Ligation via Restriction Digest of both the Expression Gene and Backbone Plasmid.
        
        Lab 11- Restriction Digest Products will be Isolated via Gel Electrophoresis. Isolated Products will then be Ligated to form a Resultant Vector that can be Transformed into Bacteria.
        
        Lab 12- Resultant Vector from Lab 11 will be Transformed into E. Coli Cells and Plated.
        
        Lab 13- A Pellet (formed via inoculation of plated cells in Lab 12) will be used to purify a larger concentration of the resultant vector from Lab 11.
    - - Lab 13- Induction Conditions for E. Coli (transformed with the Lab 11 resultant vector ) will be chosen for Lab 14.
        
        Lab 14- 14. E. Coli grown under a chosen set of conditions (Lab 13) will be lysed and put under SDS-page to determine protein composition. This will determine the favorable conditions for expression of the recombinant human LDHA in E. coli.