characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-06-01T17:02:23Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:51:27Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2235068
characteristics.characteristic: External Id
characteristics.text: ICGEB at New Delhi
characteristics.characteristic: INSDC center name
characteristics.text: 2014-06-01T17:02:23Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:51:27Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS364384
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2045:iBF 5d
characteristics.characteristic: Submitter Id
characteristics.text: 6 to 7 day
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Asian malaria mosquito
characteristics.characteristic: common name
characteristics.text: Protocols: An. stephensi from laboratory stock were reared under controlled conditions at 28 + 2degreesC, 70 -75% humidity. Adult mosquitoes were fed with 2% sterile glucose solution and water soaked raisin.Samples were prepared from An. stephensi at 5 different conditions. 5-6 days old sugar fed adult female mosquitoes were collected. Blood fed and parasite infected mosquitoes were collected at two time points, at 42 hours and five days post blood feeding. Whole body of all mosquito samples were stored in Trizol (Invitrogen) until RNA extraction. Stock solution of Plasmodium vinckei petteri 279 BY was thawed and injected into 5 weeks old BALB/C mice. When gametocytemia reached 0.05%, naive 4-5 days old female mosquitoes were fed on infected mice. Midguts of mosquitoes on 5th day post infection (dpi) were dissected, stained in mercurochrome and observed for the presence of oocysts under microscope. Presence of oocysts confirmed infection of mosquitoes by Plasmodium parasite. BF 42h and BF 5d mosquitoes were fed on normal mice blood and were collected at specified time points. Total RNA enriched in small RNA population was extracted using miRNeasy kit (Qiagen) as per the manualﾒs protocol. Quality and quantity of RNA was checked by using Agilent 2100 Bioanalyzer RNA Nano 6000 kit. Illumina Truseq small RNA libraries were made as per the manufacturer's instructions (Illumina Inc). 1ug of total RNA was ligated with 3prime and 5prime adaptors followed by reverse transcription using RT primers. Following PCR amplification of the adaptor enriched fragments, population of small RNA within length 140-160 bps were ran and eluted from 6 % TBE PAGE gel. Eluted product was precipitated using sodium acetate and ethanol and was dissolved in RNAase free water. These RNA samples were sent for high throughput sequencing using Illumina Genome Analyzer II.
characteristics.characteristic: description
characteristics.text: wild type
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0005168
characteristics.characteristic: genotype
characteristics.text: Anopheles stephensi
characteristics.characteristic: organism
characteristics.text: Anopheles stephensi
characteristics.characteristic: organism
characteristics.text: Anopheles stephensi
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000383
characteristics.characteristic: sex
characteristics.text: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: iBF 5d
characteristics.characteristic: title