characteristics.text: LIBEST_019436
characteristics.characteristic: External Id
characteristics.text: SAMN00150068
characteristics.characteristic: External Id
characteristics.text: Washington University School of Medicine
characteristics.characteristic: INSDC center name
characteristics.text: 2006-04-05T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:44:18.530Z
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: pDNR-LIB; Site_1: Sfi IB (5'-GCCTC-3'); Site_2: Sfi IA (5'-ATTAC-3'); Parasitic second-stage juveniles of G. rostochiensis were collected 11 days after inoculation of potato tubers with cysts of G.rostochiensis line 19 (avirulent) by homogenizing the whole root system and purifying juveniles by sieving and centrifugation in sucros gradient. Total RNA was isolated according to TRIzol protocol, followed by mRNA isolation (Dynabeads mRNA purification Kit, Dynal). mRNA was used in first strand cDNA synthesis according to the Creator SMART cDNA library construction protocol, followed by cDNA amplification by 22 cycles of Long-Distance (LD) PCR, SfiI digestion, cDNA size fractionation, ligation into pDNR-LIB vector and transformation into electrocompetent TOP10 E. coli. This library was contributed by Anna Tomczak and Dr. Geert Smant (Geert.Smant@wur.nl) of the Laboratory of Nematology at Wageningen University, Wageningen, Netherlands.
characteristics.characteristic: description
characteristics.text: J2 parasitic stage, 11 days post inoculation with cysts
characteristics.characteristic: dev-stage
characteristics.text: DH10B
characteristics.characteristic: lab-host
characteristics.text: Globodera rostochiensis
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_31243
characteristics.characteristic: organism
characteristics.text: SfiI B
characteristics.characteristic: re_1
characteristics.text: SfiI A
characteristics.characteristic: re_2
characteristics.text: line 19
characteristics.characteristic: strain
characteristics.text: Globodera_rostochiensis_EST
characteristics.characteristic: title
characteristics.text: plasmid
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/SO_0000155
characteristics.characteristic: v_type
characteristics.text: pDNR-LIB
characteristics.characteristic: vector