Next Article in Journal
Postsynthetic On-Column 2′ Functionalization of RNA by Convenient Versatile Method
Previous Article in Journal
Point-of-Use Rapid Detection of SARS-CoV-2: Nanotechnology-Enabled Solutions for the COVID-19 Pandemic
 
 
Article

Discrimination of CpG Methylation Status and Nucleotide Differences in Tissue Specimen DNA by Oligoribonucleotide Interference-PCR

Department of Biochemistry and Genome Biology, Hirosaki University Graduate School of Medicine, 5 Zaifu-cho, Hirosaki, Aomori 036-8562, Japan
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2020, 21(14), 5119; https://doi.org/10.3390/ijms21145119
Received: 28 May 2020 / Revised: 10 July 2020 / Accepted: 17 July 2020 / Published: 20 July 2020
(This article belongs to the Section Molecular Biology)
Oligoribonucleotide (ORN) interference-PCR (ORNi-PCR) is a method in which PCR amplification of a target sequence is inhibited in a sequence-specific manner by the hybridization of an ORN with the target sequence. Previously, we reported that ORNi-PCR could detect nucleotide mutations in DNA purified from cultured cancer cell lines or genome-edited cells. In this study, we investigated whether ORNi-PCR can discriminate nucleotide differences and CpG methylation status in damaged DNA, such as tissue specimen DNA and bisulfite-treated DNA. First, we showed that ORNi-PCR could discriminate nucleotide differences in DNA extracted from acetone-fixed paraffin-embedded rat liver specimens or formalin-fixed paraffin-embedded human specimens. Rat whole blood specimens were compatible with ORNi-PCR for the same purpose. Next, we showed that ORNi-PCR could discriminate CpG methylation status in bisulfite-treated DNA. These results demonstrate that ORNi-PCR can discriminate nucleotide differences and CpG methylation status in multiple types of DNA samples. Thus, ORNi-PCR is potentially useful in a wide range of fields, including molecular biology and medical diagnosis. View Full-Text
Keywords: ORNi-PCR; PCR; acetone-fixed paraffin-embedded (AFPE); formalin-fixed paraffin-embedded (FFPE); bisulfite; CpG methylation; polymorphism; mutation; epidermal growth factor receptor (EGFR) ORNi-PCR; PCR; acetone-fixed paraffin-embedded (AFPE); formalin-fixed paraffin-embedded (FFPE); bisulfite; CpG methylation; polymorphism; mutation; epidermal growth factor receptor (EGFR)
Show Figures

Figure 1

MDPI and ACS Style

Shimizu, T.; Fujita, T.; Fukushi, S.; Horino, Y.; Fujii, H. Discrimination of CpG Methylation Status and Nucleotide Differences in Tissue Specimen DNA by Oligoribonucleotide Interference-PCR. Int. J. Mol. Sci. 2020, 21, 5119. https://doi.org/10.3390/ijms21145119

AMA Style

Shimizu T, Fujita T, Fukushi S, Horino Y, Fujii H. Discrimination of CpG Methylation Status and Nucleotide Differences in Tissue Specimen DNA by Oligoribonucleotide Interference-PCR. International Journal of Molecular Sciences. 2020; 21(14):5119. https://doi.org/10.3390/ijms21145119

Chicago/Turabian Style

Shimizu, Takeshi, Toshitsugu Fujita, Sakie Fukushi, Yuri Horino, and Hodaka Fujii. 2020. "Discrimination of CpG Methylation Status and Nucleotide Differences in Tissue Specimen DNA by Oligoribonucleotide Interference-PCR" International Journal of Molecular Sciences 21, no. 14: 5119. https://doi.org/10.3390/ijms21145119

Find Other Styles
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Back to TopTop