Electrochemical screening for the selection of DNA aptamers
AssigneeAlfaisal University
InventorsShimaa Eissa, Mohammed Zourob, Raja Chinnappan, Ayesha Siddiqua
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Abstract
An electrochemical screening method for the selection of DNA aptamers against 11-deoxycortisol (11-DCL) using gold electrode for target immobilization is described. The gold electrode is used as solid matrix instead of the beads for SELEX. The selection steps (SELEX) are performed on the 11-DC modified electrode directly as the DNA library in the first round or the enriched DNA pools in the subsequent rounds were incubated on the electrode, then the unbound DNA is washed and the bound DNA is measured directly by square wave voltammetry. Then elution of the bound DNA is performed for further use.
Claims6
An electrochemical screening method for an aptamer, comprising:
- immobilizing a target analyte on an electrode by the following steps:
- a) coating an electrode with a cysteamine hydrochloride to create a cysteamine-modified electrode, wherein the electrode is made of a metal;
- b) washing the cysteamine-modified electrode with an ethanol to remove an unbound cysteamine hydrochloride from the cysteamine-modified electrode;
- c) adding a cross linker 1, 4-phenylene diisocyanate to an amine group of cysteamine hydrochloride on the surface of the cysteamine-modified electrode;
- d) washing with a methanol to remove any unbound cross linker 1, 4-phenylene diisocyanate from the surface of the cysteamine-modified electrode;
- and
- e) incubating the cysteamine-modified electrode in a 11-deoxycortisol solution for crosslinking the-target analyte and 11-deoxycortisol molecule on the surface of the cysteamine-modified electrode to create a 11-deoxycortisol-modified electrode to perform an electrochemical selection of an aptamer, wherein the aptamer is selected from a group consisting of a SEQ ID NO:
- 3-DC 3, SEQ ID NO:
- 4-D 28, SEQ ID NO:
- 5-DC 7, SEQ ID NO:
- 6 DC-13, SEQ ID NO:
- 7 DC-19, SEQ ID NO:
- 8 DC-21, SEQ ID NO:
- 9 DC-27, SEQ ID NO:
- 10 DC-15, SEQ ID NO:
- 11 DC-17, SEQ ID NO:
- 12 DC-23 and SEQ ID NO:
- 13 DC-14.
Dependencies are inferred from the claim text.