High throughput sequencing of 16S rRNA amplicons from individual Ixodes ricinus nymphs, collected in Poitou-charentes, France
nymphs were collected and rinsed in 1% bleach for 30 seconds, then rinsed 3 times in water for 1 minute. Individuals were crushed and total DNA was extracted using Blood and Tissue kit (Qiagen). PCR amplifications were perfomed using Q5 polymerase (NEB) with U341F and Bakt_805R primers. Amplicons were checked on 1% agarose gels, and purified using NucleoSpin Gel and PCR Cleanup kit (Macherey-Nagel). Amplicons were quantified using Qubit 2.0 fluorometer, and sent for Illumina MiSeq sequencing at the ICM facility (https://igenseq.institutducerveau-icm.org/en/).