Optimization of binding, washing and elution buffer for development of DNA isolation kit
DNA isolation is also known as the isolation of DNA from the desired cells. This procedure is one of the most crucial part in DNA analysis for scientists and they can buy any ready-to-use DNA isolation kits as there are many choices in the market. However, the chaotropic salts used in conventional k...
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| Format: | Undergraduate Final Project Report |
| Published: |
2020
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| Online Access: | http://discol.umk.edu.my/id/eprint/4149/ |
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| Summary: | DNA isolation is also known as the isolation of DNA from the desired cells. This procedure is one of the most crucial part in DNA analysis for scientists and they can buy any ready-to-use DNA isolation kits as there are many choices in the market. However, the chaotropic salts used in conventional kits for binding step inhibit the downstream process of PCR and deteriorate when exposed to air. Hence, a better and faster DNA isolation process with better performance of DNA isolation will be tested in order to replace the conventional one. Thus, this research is to produce an improved procedure of the DNA isolation that comes with an ideal formulation of binding solution, organic wash solution and elution buffer. Currently, the most recommended way to improve the performance is replacing chaotropic salts to organic solvents and followed by elution after washing step. The reason of replacing these compounds is because they prevent the inhibition of downstream process for PCR. In addition, the tested method will begin with optimization of DNA binding by using selected organic acids and salt, and it is then followed by optimization of wash step. Then, the comparison with DNA extraction kit in the market was carried out and the data collected was analysed by ANOVA test. Thus, a result of an improved process of DNA isolation with optimum formulation of binding solution, wash solution are expected. Binding buffer of glycine is showing the best concentration of DNA result among the others and the washing buffer of 90% ethanol with 10Mm NaCI and 10Mm Tris-Cl is showing the best yield of DNA ratio 260/280. |
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