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  • A Simple Mouthwash Method For Genomic DNA Isolation in Molecular Studies

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    Rosnah Binti Zain
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    This study describes a simple and inexpensive method for isolating genomic DNA from exfoliated oral cells using a mouthwash technique. Twenty-two subjects swished saline in their mouths for 60 seconds and spit into a collection tube. DNA was extracted from the samples using a saliva DNA isolation kit. The extracted DNA showed good purity and yield, with an average ratio of 1.94 for A260/A280. All samples were successfully genotyped using PCR to amplify the p53 gene, demonstrating that the isolated DNA was of high enough quality to support molecular analysis. The results indicate that this non-invasive mouthwash method provides sufficient amounts of high molecular weight DNA for downstream applications like PCR-based assays.

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    This study describes a simple and inexpensive method for isolating genomic DNA from exfoliated oral cells using a mouthwash technique. Twenty-two subjects swished saline in their mouths for 60 seconds and spit into a collection tube. DNA was extracted from the samples using a saliva DNA isolation kit. The extracted DNA showed good purity and yield, with an average ratio of 1.94 for A260/A280. All samples were successfully genotyped using PCR to amplify the p53 gene, demonstrating that the isolated DNA was of high enough quality to support molecular analysis. The results indicate that this non-invasive mouthwash method provides sufficient amounts of high molecular weight DNA for downstream applications like PCR-based assays.

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    11 views2 pages

    A Simple Mouthwash Method For Genomic DNA Isolation in Molecular Studies

    Uploaded by

    Rosnah Binti Zain
    This study describes a simple and inexpensive method for isolating genomic DNA from exfoliated oral cells using a mouthwash technique. Twenty-two subjects swished saline in their mouths for 60 seconds and spit into a collection tube. DNA was extracted from the samples using a saliva DNA isolation kit. The extracted DNA showed good purity and yield, with an average ratio of 1.94 for A260/A280. All samples were successfully genotyped using PCR to amplify the p53 gene, demonstrating that the isolated DNA was of high enough quality to support molecular analysis. The results indicate that this non-invasive mouthwash method provides sufficient amounts of high molecular weight DNA for downstream applications like PCR-based assays.

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    A simple mouthwash method for genomic DNA isolation in molecular studies

    Article type: Meeting Abstract Content: Background: Application of PCR techniques requiring only minute amounts of Genomic DNA. Thus, a less invasive, simpler to perform, and cheaper method to obtain DNA from exfoliated cells is desirable. We aim to develop a method that can obtain high quality of genomic DNA from one sample that allows for numerous application of PCR analysis. Objective: This study describes a simple, inexpensive and non invasive protocol to isolate a high quality of genomic DNA from exfoliated cells by using swish method. Methodology: Twenty two subjects vigorously swished 10 ml of normal saline in their mouth for 60 s and spitted into a collection tube. DNA extraction assay was performed by using saliva DNA isolation kit (Norgen, USA). The washed pellets were suspended in TE buffer and analyzed for the quality and purity of DNA content by using the NanoDrop Spectrophotometer. A ratio of A260/A280 was calculated. The extracted genomic DNA was amplified with primers of p53 intron 6 by using PCR machine. The presence of amplified DNA was then confirmed by electrophoreses analysis, which DNA bands were scanned by Typhoon 9410 variable imager. Results: In this study, the extracted genomic DNA demonstrated an average value of 1.94 O.D. in DNA content purity and 42.9 g/l in DNA yields. The electrophoresis images of the DNA products showed visible and detectable bands of higher molecular weight DNA in all the samples. Conclusion: The results showed that the extracted genomic DNA from the exfoliated cells by applying the swish method, that provides substantially larger amounts and higher molecular weight of DNA for down-stream DNA identification application. In addition, all samples were successfully genotyped by PCR-based assays for p53 gene intron 6 regions, which confirmed that the quality of isolated DNA was reliable in supporting the PCR amplification for the molecular studies. Author Source ISSN DOI Volume Page Year Khor, G. H. ; Zain, R. B. ; Abu Hassan, M. I. ; Ahmed, R. Khan, Hbsg ; Younis, L. T. Oral Oncology 1368-8375 10.1016/j.oos.2009.06.587 45 7 2009

    Keyword:
    Oral squamous cell carcinoma, OSCC, lichenoid lesions, lichen planus, oral cancer, oral tumours, pemphigus, traumatic eosinophilic granuloma, aphthous ulcers, oral mucosal lesions, betel chewers mucosa, betel quid related lesions, betel quid, areca quid, tobacco quid, oral cancer screening, training and calibration, early detection, oral cancer awareness, biobanking, tissue bank, databank, oral cancer, tissue bank, research credibility, research ethics.

    Please Cite As: KHOR, G. H., ZAIN, R. B., ABU HASSAN, M. I., AHMED, R., KHAN, H. & YOUNIS, L. T. 2009. A simple mouthwash method for genomic DNA isolation in molecular studies. Oral Oncology, 221-221. URL: http://apps.webofknowledge.com/full_record.do?product=WOS&search_mode=M arkedList&qid=12&SID=4EcD1HbJnNn6@H@k7ME&page=1&doc=3&colName= WOS http://www.sciencedirect.com/science/article/pii/S1744789509005884

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