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Development of an Ammonium Sulfate DNA Extraction Method for Obtaining Amplifiable DNA in a Small Number of Cells and Its Application to Clinical Specimens
Joint Authors
Oh, Seo Young
Kim, Wook Youn
Hwang, Tae Sook
Han, Hye Seung
Lim, So Dug
Kim, Wan Seop
Source
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-04-17
Country of Publication
Egypt
No. of Pages
10
Main Subjects
Abstract EN
DNA extraction from microdissected cells has become essential for handling clinical specimens with advances in molecular pathology.
Conventional methods have limitations for extracting amplifiable DNA from specimens containing a small number of cells.
We developed an ammonium sulfate DNA extraction method (A) and compared it with two other methods (B and C).
DNA quality and quantity, β-globin amplification, and detectability of two cancer associated gene mutations were evaluated.
Method A showed the best DNA yield, particularly when the cell number was very low.
Amplification of the β-globin gene using DNA from the SNU 790 cell line and papillary thyroid carcinoma (PTC) cells extracted with Method A demonstrated the strongest band.
BRAFV600E mutation analysis using ethanol-fixed PTC cells from a patient demonstrated both a “T” peak increase and an adjacent “A” peak decrease when 25 and 50 cells were extracted, whereas mutant peaks were too low to be analyzed using the other two methods.
EGFR mutation analysis using formalin-fixed paraffin-embedded lung cancer tissues demonstrated a mutant peak with Method A, whereas the mutant peak was undetectable with Methods B or C.
Method A yielded the best DNA quantity and quality with outstanding efficiency, particularly when paucicellular specimens were used.
American Psychological Association (APA)
Oh, Seo Young& Kim, Wook Youn& Hwang, Tae Sook& Han, Hye Seung& Lim, So Dug& Kim, Wan Seop. 2013. Development of an Ammonium Sulfate DNA Extraction Method for Obtaining Amplifiable DNA in a Small Number of Cells and Its Application to Clinical Specimens. BioMed Research International،Vol. 2013, no. 2013, pp.1-10.
https://search.emarefa.net/detail/BIM-1004574
Modern Language Association (MLA)
Oh, Seo Young…[et al.]. Development of an Ammonium Sulfate DNA Extraction Method for Obtaining Amplifiable DNA in a Small Number of Cells and Its Application to Clinical Specimens. BioMed Research International No. 2013 (2013), pp.1-10.
https://search.emarefa.net/detail/BIM-1004574
American Medical Association (AMA)
Oh, Seo Young& Kim, Wook Youn& Hwang, Tae Sook& Han, Hye Seung& Lim, So Dug& Kim, Wan Seop. Development of an Ammonium Sulfate DNA Extraction Method for Obtaining Amplifiable DNA in a Small Number of Cells and Its Application to Clinical Specimens. BioMed Research International. 2013. Vol. 2013, no. 2013, pp.1-10.
https://search.emarefa.net/detail/BIM-1004574
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1004574