characteristics.text: LIBEST_023752
characteristics.characteristic: External Id
characteristics.text: SAMN00166711
characteristics.characteristic: External Id
characteristics.text: University of Maine
characteristics.characteristic: INSDC center name
characteristics.text: 2009-10-10T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2015-05-20T14:43:54.173Z
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: In the presence of sprouted potato tubers, mycelial cultures of quinate- induced RHS 1AP and uninduced RHS 1AP were grown and poly(A)+ RNA was extracted. To prepare subtracted cDNAS, we used the BD Clontech PCR-SELECT cDNA subtraction kit (BD Clontech). Normalization of cDNA was checked using the BD Advantage 2 PCR enzyme kit (BD Biosciences Clontech) and primers specific for the R. solani (AG3) glyceraldehyde-3-phosphate dehydrogenase (GPDH) (GenBank AF339929) or the R. solani (AG 3) specific -tubulin gene sequence, provided by Dolores Gonz Lez (Laboratorio de Sistem Tica Molecular Instituto de Ecolog A, A. C., Mexico). cDNA from quinate-induced RHS 1AP was subtracted with excess CDNA from untreated RHS 1AP and was checked for normalization of the two constitutive genes, GPDH and -tubulin. In the case of the GPDH gene, the respective pcr product became slightly visible after 31 cycles of subtracted cDNA amplification, whereas copious PCR product was generated after 19 cycles of unsubtracted cDNA amplification. Similarly, the -tubulin gene PCR product became visible following 33 cycles of subtracted cDNA amplification, but after only 18 cycles of unsubtracted cDNA amplification. Subtracted cDNAS were cloned into a T/A-based vector (TOPO PCR cloning kit - Invitrogen). The Eppendorf Fastplasmid mini kit (Brinkmann) was used for plasmid purification, and DNA sequencing was conducted using an ABI3700 capillary DNA sequencer.
characteristics.characteristic: description
characteristics.text: Rhizoctonia solani
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_456999
characteristics.characteristic: organism
characteristics.text: RHS 1AP (AG 3)
characteristics.characteristic: strain
characteristics.text: Quinate-induced cDNA library of Rhizoctonia solani
characteristics.characteristic: title