organism: Anopheles stephensi
description: 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. 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.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2014-06-01T17:02:23Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T16:51:33Z
attributes.tag: External Id
attributes.value: SAMEA2235069
attributes.tag: INSDC center name
attributes.value: ICGEB at New Delhi
attributes.tag: INSDC first public
attributes.value: 2014-06-01T17:02:23Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T16:51:33Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2045:SF
attributes.tag: age
attributes.value: 6 to 7 day
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: Asian malaria mosquito
attributes.tag: genotype
attributes.value: wild type
attributes.tag: sample name
attributes.value: E-MTAB-2045:SF
attributes.tag: scientific_name
attributes.value: Anopheles stephensi
attributes.tag: sex
attributes.value: female
attributes.tag: specimen with known storage state
attributes.value: fresh specimen