DNA extraction optimization of stool from colorectal patient

Wardoyo, Eustachius Hagni and Sabrina, Yunita and Suryani, Dewi and Indriawan, Ramses DNA extraction optimization of stool from colorectal patient. Seminar Nasiona; Saintek Universitas Mataram 2019. (Unpublished)

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Abstract

Background: Diarrheal episodes of colorectal patient is often and sometimes persist, due to clinical consequences, adverse event after chemo/radio-therapy or the presence of infection. The condition led to dehydration to death. Unformed stool may change stool composition of bacterial cells, molecular inhibitor and others that differ from formed stool. This led to altered DNA concentration that altered DNA extraction protocol differ from standar protocol. Aim: to obtain optimal condition of DNA extraction protocol from unformed stool of colorectal patient Methods: DNA extraction kit used was Quick-DNA™ Fecal/Soil Microbe Miniprep Kit (Zymoresearch). There are 3 protocol done for the study, which one of it is standar protocol from manufacturer, 2nd and 3rd was modification with following principles: 1. Mechanical disruption to separate fecal material from bacterial cell, 2. Modification in speed and duration of spin during DNA binding and 3. Modification of buffer DNA elution volume. Standar protocol is applied to a matched control stool and sample stool, while 2nd and 3rd protocol is applied only to sample stool. Absorbance ratio of A260/280 ≥ 1.8 using Nanodrop™ was applied to final DNA reflecting DNA purity. It was measured three times and the average value is reported as a final value. Result: The absorbance ratio of A260/280 ≥1.8 was found in 3rd protocol using fecal sample from the patient. A matched control stool and sample stool in standar protocol was 1,9 and 1,78, respectively. Second protocol 1.79 and third 1.81. Conclusion: Third protocol developed as the optimal DNA extraction for unformed stool from colorectal patient in chemotherapy.

Item Type: Article
Keywords (Kata Kunci): DNA extraction to unformed stool, colorectal patient
Subjects: R Medicine > RZ Other systems of medicine
Divisions: Fakultas Kedokteran
Depositing User: dr.Sp.MK Eustachius Hagni Wardoyo
Date Deposited: 03 Dec 2019 01:13
Last Modified: 03 Dec 2019 01:13
URI: http://eprints.unram.ac.id/id/eprint/15088

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