Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 Aug 19;4:988. doi: 10.1038/s42003-021-02503-5

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Fig. 4 — a A schematic diagram describing blocking RPA with MBD2 protein as a blocking agent (MBDi-RPA). MBD proteins bind to any methylated CpG sites in a target sequence, resulting in the suppression of amplification. MBDi-RPA can be used to discriminate CpG methylation status. b A target sequence in the human CDKN2A (p14ARF) gene in HCT116. Forward and reverse primer positions are highlighted in blue and green, respectively. c Results of MBDi-RPA. M, molecular weight marker. Amplified CDKN2A (p14ARF) is indicated by an arrowhead. d Results of DNA sequencing analysis. RPA or MBDi-RPA amplicons from c were sequenced using a reverse primer. e A schematic diagram describing MBDi-RPA of in vitro CpG-methylated plasmid DNA. The CDKN2A (p14ARF) sequences shown in b were cloned into plasmid vectors. The sequence corresponding to the Gx4 allele was subsequently methylated in vitro. f Results of MBDi-RPA from individual Gx4 and Gx5 plasmids or a plasmid mixture. g Results of DNA sequencing analysis. RPA and MBDi-RPA amplicons shown in f (plasmid mixture) were sequenced using a reverse primer.