Sequencing and Alignment
PCR products were kept in PCR plates sealed with adhesive film or caps
and sent oversea via French post services to be sequenced. Sanger
sequencing was handled by Microsynth facility in Lyon, France. Forward
and reverse sequencing were realized with the same set of primers than
for the PCR reaction. Fragment sequences were aligned using BLAST (NCBI,
NIH) against the standard databases (nucleotide collection nr/nt)[40,41] and Anopheles species identifications were
retrieved for forward and reverse sequences individually and integrated
in our data and metadata files.
Reference sequences of ITS2 region of 14 Anopheles species found
in French Guiana are available online on the NIH website (PopSet:
870902931) [31] .