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 5;11:15869. doi: 10.1038/s41598-021-95563-w

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Dual molecular DNA barcoding and PCR-driven concatemerization applied to SARS-CoV-2 diagnostics. (A) Reverse transcription is performed with oligonucleotides targeting a Covid-19 region, but in addition containing a unique molecular barcode per sample and a common adapter sequence (Gibson). Reverse transcription assays (cDNA) performed for multiple candidates are pooled together for amplification. PCR amplification is performed with oligonucleotides targeting the common Gibson sequence and a Covid-19-specific primer hosting a second molecular barcode as well as a complementary Gibson sequence. During PCR cycles, amplified products are able to concatenate thanks to the Gibson flanking sequences. We have called this particular PCR strategy “Concat-PCR”. (B) DNA electropherogram (Agilent TapeStation system) illustrating concatemer products obtained with PCR primers at 1uM final concentration. (C) DNA electropherogram for a concat-PCR assay performed with PCR primers at 0.1uM final concentration, demonstrating a preferential production of long concatemers in contrast to those observed in (B). (D) Timeline illustrating the various molecular biology steps required for samples preparation prior engaging samples into MinION DNA sequencing.