Sample 14
Anopheles coluzzii
Protocols: 12 mosquitoes per pool, in each of two biological replicate pools, were collected 3 d after infection by oral feeding with ONNV and homogenized in TriReagent (Sigma). The A. coluzzii Ngousso strain was originally initiated with mosquitoes collected in Cameroon in January 2006, and belongs to the M molecular and Forest chromosomal forms (Harris et al., 2010). Mosquitoes were reared under standard conditions at 26°C and 80% relative humidity, with a 12 h light/dark cycle as previously described (Mitri et al., 2015). Double-stranded RNAs were synthesized from PCR amplicons using the T7 Megascript Kit (Ambion) as described (Carissimo et al., 2015). For each targeted gene, 500 ng of dsRNA was injected using glass capillary needles into the thorax of cold-anesthetized 1-2 d-old A. coluzzii females using a nano-injector (Nanoject II, Drummond Scientific). The 10- to 30-nt RNA fragments were purified from the small RNA fraction (<200nt) on a 15% urea PAGE (BioRad). Small RNA libraries were then prepared using TruSeq Small RNA Library reagents (Illumina). 15-30nt RNA extracted from polyacrylamide gel were ligated with 3'- and 5'- TruSeq adapters, as described in the original protocol. Synthesis of cDNA was performed by reverse transcription. The cDNA product was then specifically amplified by 11 cycles of PCR and products purified on Agencourt AMPure XP beads (Beckman Coulter Genomics, # A63881).
ENA first public
2018-06-28
ENA last update
2018-01-31
External Id
SAMEA104557098
INSDC center alias
IP
INSDC center name
Institut Pasteur
INSDC first public
2018-06-28T17:03:06Z
INSDC last update
2018-01-31T14:11:36Z
INSDC status
public
Submitter Id
E-MTAB-6444:Sample 14
broker name
ArrayExpress
organism part
whole organism
replicate
2
sample name
E-MTAB-6444:Sample 14
sex
female
strain
M