characteristics.text: LIBEST_019640
characteristics.characteristic: External Id
characteristics.text: SAMN00150267
characteristics.characteristic: External Id
characteristics.text: Institute of Animal Sciences, Chinese Academy of Agricultural Sciences
characteristics.characteristic: INSDC center name
characteristics.text: 2006-05-08T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:44:23.817Z
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 total RNA was isolated from the spleen samples using TRIZOL Kit according to the manufacturer's protocol and digested with RNase-free DNase I to eliminate chromosomal DNA. The concentration of RNA was determined by spectrophotometer. A RNA pool containing equal amounts RNA of 20 chickens samples was established. Each RNA pool was reverse transcribed with M-MLV reverse transcriptase using one of the three anchor primers. The cDNA of the two samples were amplified with random primer and corresponding anchor primer. For the DDRT-PCR combinations of the 3 anchor primers together with 18 random primers. The DNA bands were separated on a polyacrylanlid gels and identified by silver staining. The differentially expressed bands were extracted from the gel and reamplified using the same primers set previously. The PCR products were identified by reverse Northern dot bloting with the DIG-labeled cDNA using DIG High Prime DNA Labeling and Detection Kit according to the manufacturer's protoco1. The positive DNA fragments were cloned into pGEM-T easy vector and then transformed into E.coli DH5a competent cells. After identification by enzyme digestion, the positive clones from the ampicillin positive LB plate were selected and sequenced. Uncertain fragment were identified by semiquantitative RT-PCR.
characteristics.characteristic: description
characteristics.text: spleen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0002106
characteristics.characteristic: organ
characteristics.text: Gallus gallus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_9031
characteristics.characteristic: organism
characteristics.text: spleen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0002106
characteristics.characteristic: tissue-type
characteristics.text: ARBOR ACRES spleen
characteristics.characteristic: title
characteristics.text: pGEM-T easy
characteristics.characteristic: vector