characteristics.text: LIBEST_022922
characteristics.characteristic: External Id
characteristics.text: SAMN00165468
characteristics.characteristic: External Id
characteristics.text: University of Bologna
characteristics.characteristic: INSDC center name
characteristics.text: 2008-10-31T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:45:59.220Z
characteristics.characteristic: INSDC last update
characteristics.text: live
characteristics.characteristic: INSDC status
characteristics.text: Generic
characteristics.characteristic: NCBI submission model
characteristics.text: Generic.1.0
characteristics.characteristic: NCBI submission package
characteristics.text: HcrVf2 trangenic Gala
characteristics.characteristic: cultivar
characteristics.text: Total RNA was extracted from frozen pooled leaf tissues of seven independent scab-resistant Gala lines transformed with the HcrVf2 gene (Belfanti et al., 2004) and the scab susceptible cultivar Gala challenged with Venturia inaequalis. Poly(A)+mRNA was isolated from total RNA using the FastTrack 2.0 kit (Invitrogen). The library was prepared using the PCR-Select Subtractive Hybridization Kit (Clontech). A 'forward' subtraction was performed to isolate fragments corresponding to genes whose expression level is increased in resistant genotypes during interaction with the pathogen. This was done by subtracting the cDNA prepared from the susceptible cv. Gala pooled leaves collected just after inoculum (used as 'driver') to the cDNA from Ga2 resistant pooled leaves collected at various times after inoculum (used as 'tester'). A 'reverse' subtraction was also carried out to isolate fragments corresponding to genes whose expression level is decreased following the onset of plant/pathogen interaction. Each type of subtraction was performed twice, starting from total RNA obtained during independent extractions. The ligated cDNA fragments were transformed into E.coli DH5 alpha host cells.
characteristics.characteristic: description
characteristics.text: E. coli DH5alpha
characteristics.characteristic: lab-host
characteristics.text: Malus domestica
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_3750
characteristics.characteristic: organism
characteristics.text: young leaves
characteristics.characteristic: tissue-type
characteristics.text: Venturia inaequalis challenged cDNA suppression subtractive hybridization library
characteristics.characteristic: title
characteristics.text: Plasmid
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/SO_0000155
characteristics.characteristic: v_type
characteristics.text: pGEM-T Easy
characteristics.characteristic: vector