Abi prism 3100 capillary automatic dna analyzer

The genetic characterization of the RT-LAMP amplicons from some positive samples from human serum obtained patient infected with ZIKV was executed by the Sanger sequencing method as previously described^{45 (link)}. RT-LAMP amplicons were directly purified using Gel Band Purification Kit (GE) and illustra GFX PCR DNA according to the manufacturer’s protocol and eluted in 30 μL of water. Purified RT-LAMP fragments were directly sequenced using the primer FIP and the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems, USA) according to the manufacturer´s instructions and run on an ABI Prism 3100 Capillary Automatic DNA Analyzer. After sequencing by the Sanger method, the sequences were analyzed using the Bioedit software, v7.0.5 and submitted to NCBI BLAST database (http://www.ncbi.nlm.nih.gov/blast/Blast.cgi) to find the most closely ZIKV strain.

The genetic characterization of the LAMP fragments from two field positives samples from A. aegypti and C. quinquefasciatus was performed by the Sanger sequencing method. Amplicons from RT-LAMP reaction were directly purified using illustra GFX PCR DNA and Gel Band Purification Kit (GE) according to the manufacturer´s instructions and eluted in 30 μL of water. Purified amplicons were directly sequenced using the primer FIP and the BigDye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems,USA) as established by the manufacturer and run on an ABI Prism 3100 Capillary Automatic DNA Analyzer. Sequences of fragments were analyzed using the Bioedit software, v7.0.5 and submitted to NCBI BLAST database (http://www.ncbi.nlm.nih.gov/blast/Blast.cgi) to identify the most closely ZIKV strain.