Competition Dialysis in Biochemistry: Discovery of Ligands

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Introduction

Competition dialysis is a new tool that is considered very powerful for the discovery of ligands.
Such ligands binds to the nucleic acid with sequence selectivity. The sequence selectivity is also
known as structural connectivity. This particular method is based on a very firm principle of
thermodynamic which makes its implementation very simple. In the experiment of competition
dialysis, arrays of the sequence and nucleic acid is dialyzed against a common test ligand
solution(Bewley and Black 2012).
1.How does Brad chaires do the competition dialysis?
This was a procedural process. I t started with the Preparation of the solution of nucleic acid and
dialysate solution. The first and the second generation of the assays utilized 200mL of 1uM
solution. This was introduced into the unit of dialysis with 0.5mL out of 0.75uM of the sample of
the nucleic acid. The upper limit of the concentration was found to be at 5uM.The optimal
concentration that was obtained was1-2uM.The assembly and loading of the dialysis unit took
almost two hours. The solutions of the ligand was mixed with the acid in order to establish the
equilibrium. The establishment of the equilibrium lasted for over 15 hours as the incubation
period. The concentration of the recovered samples was determined using absorbance(Davies
2013).
2. What concentration does he use? Explain and why?
The first and the second generation of the assays utilized 200mL of 1uM solution. This was
introduced into the unit of dialysis with 0.5mL out of 0.75uM of the sample of the nucleic acid.
The final concentration of 75uM was preferred since it could be stored up to a period of 6
months at a temperature of 4 degrees Celsius.
3.Why does he add surfactant? Explain and if there is a reason why?