Simultaneous Nucleic Acids Separation and Extraction Using Silica Gel-Agarose Matrix Gel

Background and Objective: Amplified product of desired gene and a digested product of DNA cloning vectors using restriction enzymes is proceeding for identification of molecular size, and restriction fragments need to be separated in agarose gel electrophoresis system, both of these steps are time-consuming and expensive procedures in molecular cloning techniques. To perform agarose gel electrophoresis, expensive agarose powder will be required. The extraction of DNA molecules from the gel is a multistep process and expensive kits are required. Materials and Methods: Based on the outcomes of the present study, a new matrix was proposed in which DNA molecules can be separated and desired fragments can be extracted and purified without the use of any commercial kit available on the market. For matrix mixture preparation, 0.5% each of agarose and silica gel was taken in 1 x TAE buffer and the agarose and silica mixture was dissolved at boiling temperature. After dissolving substances, 2 L of Ethidium Bromide (EtBr) of concentration 10 mg mLG 1 was added and solidified into a gel casting tray. The amplified PCR product and vector digested product was carried out as per the standard procedure. From the gel desired fragment of DNA was extracted. Results: The agarose and silica-formed gel are a little clear and bands also seems to be prominent. Recovery of DNA bands was almost 90%. The purity of extracted DNA is within the range. The obtained DNA is suitable for restriction digestion, ligation, PCR, sequencing, transformation, etc. Conclusion: The preparation of silica-based gel is very economical and the extraction of DNA from these gels is very simple without the utilization of any commercial gel extraction kits. Its cost is almost one-tenth of the traditional system, and a single step is required for electrophoresis and gel extraction.