characteristics.text: SAMN00165412
characteristics.characteristic: External Id
characteristics.text: LIBEST_022868
characteristics.characteristic: External Id
characteristics.text: Huazhong Agricultural University
characteristics.characteristic: INSDC center name
characteristics.text: 2008-03-24T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:45:57.653Z
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: Early flowering trifoliate orange mutant (precocious trifoliate orange)
characteristics.characteristic: cultivar
characteristics.text: The subtracted library was obtained by using cDNAs from the adult mRNAs as driver and cDNAs from juvenile mRNAs as tester. Adult and juvenile precocious trifoliate orange samples were collected in the experiment fields of National Citrus Breeding Center in Huazhong Agriculture University. Several plant organs (lateral buds, terminal buds, and shoot meristems) at different developmental stages were sampled and immediately frozen in liquid nitrogen, and then stored at 80oC until use. Adult precocious trifoliate orange have three main flushes of growth. The main one occurs in late winter or early spring (spring flush), which were sprouted and developed from a brunch of a tree in March. In 2005, young buds in the axils of leaves (lateral buds) and terminal buds of spring flushes were collected at equivalent branch positions in May, July, September, November (dormancy period), respectively. In 2006, swelling buds of last year spring flushes were collected in January and March. The juvenile trees are from potted seedlings and grown under field conditions. In January of 2005, the seeds of precocious trifoliate orange were planted in 20 cm pots containing potting mix of a commercial medium and perlite at a ratio of 3:1, the juvenile trees were watered regularly with nutrient solution. Shoot meristems of the juvenile trees were collected in March, June, September, and December, respectively. To prepare a representative sample of total RNA from the adult and juvenile precocious trifoliate orange in SSH process, different developmental stages of plant organs from roughly equal numbers of the adult and juvenile precocious trifoliate oranges were pooled, respectively. SSH library was performed using PCR Select cDNA Subtraction kit (Clontech, USA) according to the manufacturer's instructions. The final PCR products were purified using QIAquick PCR purification kit (Qiagen). The fragments obtained were cloned into the pGEM-T vector (Promega, USA) and used for the transformation of Escherichia coli DH5a. Individual clones from the library were randomly picked and stored in 384-well plates. A total of 3840 clones were collected in the forward subtracted and the reverse subtracted library. The inserts were amplified with the solution of 1ul single clone bacterial culture as PCR template, 0.5ul primer 1 (10?mol/ul), 0.5ul primer 2R (10umol/ul), 0.25 ul Taq DNA polymerase (Shanghai Sangon, China), 2.5 ul 10 x DNA polymerase buffer, 0.5 ul dNTPs (10mmo/ul) and 1.5 ul MgCl2 (25 mmol/ul), and added ddH2o to 25 ul. The PCR program is: 94 oC for 5 min; 94 oC for 30 s, 58 oC for 35s, 72 oC for 1.5 min, 30 cycles followed by a 5 min final extension at 72 oC.
characteristics.characteristic: description
characteristics.text: Juvenile
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UBERON_0000112
characteristics.characteristic: dev-stage
characteristics.text: Escherichia coli DH5alpha
characteristics.characteristic: lab-host
characteristics.text: Citrus trifoliata
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_37690
characteristics.characteristic: organism
characteristics.text: Shoot meristem
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PO_0006079
characteristics.characteristic: tissue-type
characteristics.text: Juvenile adult early flowering trifoliate orange mutant subtracted cDNA library
characteristics.characteristic: title
characteristics.text: PGEM-T
characteristics.characteristic: vector