characteristics.text: LIBEST_023281
characteristics.characteristic: External Id
characteristics.text: SAMN00165854
characteristics.characteristic: External Id
characteristics.text: Forest protection of Nanjing Forestry University
characteristics.characteristic: INSDC center name
characteristics.text: 2010-12-31T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:46:10.047Z
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: The SSH was performed with cDNA from Bursaphelenchus mucronatus strain AN5#F7-1 as the tester and B. xylophilus AN19#F1-1 as the driver. Tester and driver cDNA were digested with Rsa I, the digested tester cDNA was divided into two sub- populations and each one was ligated with one of the two adaptors provided (adaptor 1 and adaptor 2R), followed by heat denaturation of both tester and driver cDNA populations. Two hybridizations were performed to generate templates for PCR amplification. An excess of driver was added to each sample of the tester for the first hybridization reaction, leading to equalization and enrichment of differentially expressed sequences. Products of each tester sub- population (adaptor 1 and adaptor 2R) from the first hybridization were mixed and immediately subjected to a second hybridization in which fresh denatured driver cDNA was added to further enrich the differential expressed sequences. Differential expressed sequences were selectively amplified by two rounds of PCR using primers specific to the adaptors, resulting in suppression of sequences common to the two cDNA populations and exponential amplification of the differentially expressed sequences. The subtracted PCR products were purified and then cloned into the pGEM-T easy vector. The ligated DNA was transformed into Escherichia coli JM109 competent cells. Clones were plated on LB medium supplemented with X-gal, IPTG and ampicillin. Individual recombinant white clones were picked and grown in LB medium containing ampicillin on 96-well plates to get the SSH library of B. mucronatus especially expressed.
characteristics.characteristic: description
characteristics.text: Mixed stage
characteristics.characteristic: dev-stage
characteristics.text: AN5#F7-1
characteristics.characteristic: isolate
characteristics.text: JM109
characteristics.characteristic: lab-host
characteristics.text: Bursaphelenchus xylophilus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_6326
characteristics.characteristic: organism
characteristics.text: SSH cDNA library of Bursaphelenchus mucronatus especially expressed
characteristics.characteristic: title
characteristics.text: pGEM-T easy vector
characteristics.characteristic: vector