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

. 2018 Nov 21;8:17195. doi: 10.1038/s41598-018-35479-0

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

© The Author(s) 2018

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

Discrimination of a single-nucleotide difference in the EGFR locus by Two-Step ORNi-PCR. (A) Nucleotide mutation leading to a Leu858Arg amino acid change in the human EGFR locus in NCI-H1975 cells. Forward DNA sequences around the Leu858 position (underlined) of both alleles are shown. (B) Target position of ORN_EGFR_L858, a 21 base ORN targeting the human EGFR locus. The forward DNA sequence around the Leu858 position (underlined) of one allele is shown. (C) Conditions for Two-Step ORNi-PCR using ORN_EGFR_L858. (D) Results of Two-Step ORNi-PCR. M, molecular weight markers. (E) DNA sequencing of Two-Step ORNi-PCR products from (D) purified from agarose gels and subjected to DNA sequencing analysis using a forward primer. (F) Mode of discrimination of a single-nucleotide difference by Two-Step ORNi-PCR. ORN_EGFR_L858 hybridises with the intact target site but not the mutated site at an annealing plus elongation step temperature of 59 °C.