RNA-seq of Ixodes ricinus: 68_3d_MG_S292
In order to study differentially expressed genes when ticks are fed on nave or already exposed to ticks rabbits, we hatched a generation of pathogen free sibling ticks and let them grow to adult females. The latest were fed on a SPF guinea pig until fully engorged and let to lay eggs. Molted larvae were fed on SPF guinea pigs (GP) (ca 100 larvae per GP and obtained Adult female ticks, coming from the same mother, were pre-mated with sibling tick males, and used for feeding and tissue isolation. The use of individuals originated from the same female tick should decrease the variability between the samples. Two rabbits were exposed each to 25 female and 25 male ticks. Five females were collected at time points 12, 24, 48, 72 and 96 hours post exposure. Ticks were dissected and their Salivary glands (SG), Midgut (MG) and ovaries (OVA) were stored in 200L RNA latter at 4C overnight followed by at -80C. Blood rabbit samples were collected at time points 0, 12, 48, 72 and 96h. Blood was kept at 4C for 12 hours, centrifuged at 10000g for 10 min. Blood serum was collected and glycerol (10% )and thimerosal (0.01% ) were added. Samples were stored at -20C. Two weeks after the end of first rabbit exposure to ticks; a second exposure was carried out following the same protocol. Five SG samples from each time point and 3 samples of MG from each time point were sequenced.