characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2014-01-27T17:00:58Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2014-01-24T16:04:35Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2338038
characteristics.characteristic: External Id
characteristics.text: Indian Agricultural Research Institute
characteristics.characteristic: INSDC center name
characteristics.text: 2014-01-27T17:00:58Z
characteristics.characteristic: INSDC first public
characteristics.text: 2014-01-24T16:04:35Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS396268
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-2221:Juvenile
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: Cysts of H. avenae were collected from infested wheat roots growing in the plots of the Indian Agricultural Research Institute, New Delhi, India. Genetic homogeneity of the population was determined by PCR-RFLP and sequencing of ITS (Internal Transcribed Spacer) regions of rDNA (ribosomal DNA). A genetically homogenous population was then multiplied on wheat plants grown in pots kept in a greenhouse (PMID:23390348). Newly formed cysts were collected manually from the wheat roots using a dissecting microscope and kept for hatching. Hatched J2s were collected and later used for RNA extraction. Likewise, hatched J2s were also used to infect wheat roots and feeding females (FF) that developed and protruded from roots 6 to 8 weeks after infection were collected, frozen (-80 degrees C) for later RNA extraction. Total RNA was extracted from the frozen J2s and FFs using TRIzol (Invitrogen Life Technologies) according to the manufacturer's instructions. Extracted RNA was assessed for quality and quantity using an Agilent 2100 Bioanalyzer (Agilent Technologies). RNA with an RNA integrity number (RIN) of 8.0 was used for mRNA purification. mRNA was purified from 1 ug of intact total RNA using oligodT beads (TruSeq RNA Sample Preparation Kit, Illumina). The purified mRNA was fragmented at elevated temperature (90 degrees C) in the presence of divalent cations and reverse transcribed with Superscript II Reverse Transcriptase (Invitrogen Life Technologies) by priming with random hexamers. Second strand cDNA was synthesized in the presence of DNA polymerase I and RNaseH. The cDNA was cleaned using Agencourt Ampure XP SPRI beads (Beckman Coulter).
characteristics.characteristic: description
characteristics.text: juvenile
characteristics.characteristic: developmental stage
characteristics.text: Heterodera avenae
characteristics.characteristic: organism
characteristics.text: Heterodera avenae
characteristics.characteristic: organism
characteristics.text: pre-parasitic
characteristics.characteristic: phenotype
characteristics.text: Heterodera avenae
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PATO_0000384
characteristics.characteristic: sex
characteristics.text: Juvenile
characteristics.characteristic: title