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. 2018 Jan 8;8:86. doi: 10.1038/s41598-017-18216-x

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© 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

Nested/PCR-RFLP of P. vivax DNA samples. 15 DNA samples from P. vivax infected individuals from different parts of Amazon: Porto Velho/Rondônia State, Brazil (PvPV/RO1 and 2), Guyana (PvGuy), Ariquemes/Rondônia State, Brazil (PvAri/RO), Venezuela (PvVen), French Guiana (PvFrGui), Novo progresso/Pará State, Brazil (PvNP/PA), Rio Pardo/Amazonas State, Brazil (PvRP/AM1, 2 and 3), Humaita/Amazonas State, Brazil (PvHu/Am1, 2, 3 and 4) and unknown city in Amazonas State, Brazil (PvAM) were used for Nested PCR amplification followed by digestion with HpyCH4III restriction enzyme. 3% Agarose gel stained with ethidium bromide. Name tags above gels indicated the patients according to Additional file 2. Reactions were performed simultaneously in the same thermocycler and splited in different gels. MM: 1 kb Plus Ladder. D: Digested (8 μL of digestion); ND: Non Digested (the equivalent amount of PCR product used in the digestion, 6.5 μL of samples and 5 μL of controls); PC Pv: positive control of P. vivax (pool of samples from infected patient from Amazonia); PC Ps: positive control of P. simium (Alouatta g. clamitans infected with P. simium previously sequenced³⁰); NC: negative Control.