characteristics.text: SAMN00164646
characteristics.characteristic: External Id
characteristics.text: LIBEST_022408
characteristics.characteristic: External Id
characteristics.text: Institute of Botany,Chinese Academy of Sciences
characteristics.characteristic: INSDC center name
characteristics.text: 2007-12-28T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:45:44.227Z
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: callus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PO_0005052
characteristics.characteristic: cell type
characteristics.text: Saussurea medusa cell cultures are cultured in Murashige and Skoog (1962) supplemented with 3% sucrose, 2 mg/l C1 NAA and 0.5 mg/l C1 6-BA 1% agar under control conditions as well as subjected to cold (0-4C), stress for 3, 6, 9, 12 d. cDNA Library was constructed using total RNA from cold stressed cell cultures. Total RNAs from different treatments were extracted and equal amounts were pooled before mRNA isolation. First cDNA synthesis was conducted by coverting poly(A)+RNA to double - stranded cDNA by using primer: 5'-ATTCTAGAGGCCGAGGCGGCCGACATG-d(T)30N C1N-3'(N = A, G, C, or T; N C1 = A, G, or C)and 5'-AAGCAGTGGTATCAACGCAGAGTGGCCATTACGGCCGGG-3'. Double stranded cDNAs were amplified LD PCR with primers containing SfiIA and SfiIB. The purified PCR products, representing the entire cloned cDNA population, were used as a driver for normalization. The normalized cDNAs were digested and then cloned into SfiIA and SfiIB digested pDNR-LIB plasmid vector (Clontech). The primary library had a titer of 2.5 x 106 pfu/ml in which 98% clones were recombinants and the insert cDNAs were from 0.5 kb to 3 kb in length. Screening of the cDNA Library by PCR using primers of M13F:5'-GTAAAACGACGGCCAGT-3' and M13R:5'-AAACAGCTATGACCATGTTCA-3'.
characteristics.characteristic: description
characteristics.text: cell cultures of 3,6,9,12 days
characteristics.characteristic: dev-stage
characteristics.text: Saussurea medusa
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_137893
characteristics.characteristic: organism
characteristics.text: SfiIA
characteristics.characteristic: re_1
characteristics.text: SfiIB
characteristics.characteristic: re_2
characteristics.text: Cell cultures of Saussurea medusa
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