characteristics.text: SAMN00152647
characteristics.characteristic: External Id
characteristics.text: LIBEST_020876
characteristics.characteristic: External Id
characteristics.text: Sri Krishnadevaraya University
characteristics.characteristic: INSDC center name
characteristics.text: 2007-02-27T00:00:00Z
characteristics.characteristic: INSDC first public
characteristics.text: 2017-10-11T17:45:02.980Z
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: A-1
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0006268
characteristics.characteristic: cultivar
characteristics.text: Seeds were surface sterilized with 0.1% (w/v) sodium hypo chlorite solution for 5 min, thoroughly rinsed with distilled water and germinated in plastic pots contained soil and sand (2:1) mixture and allowed to grow under natural photoperiod (10-12 hrs, 28 +/- 4 o C) in the departmental botanic garden. Seedlings were maintained at 100% soil moisture levels (SMLs) by watering daily. Nineteen day-old plants were subjected to moisture stress by with holding water. Leaf samples collected from unstressed (100% SMLs) and stressed (25% SMLs) plants, flash frozen and stored at -80oC. The frozen samples were used for RNA extraction (LiCl2 method). Poly(A) mRNA was isolated using streptavidin paramagnetic particles (Roche, Germany). First-strand cDNA synthesis was initiated from the enriched mRNA of driver (unstressed) using biotin labeled poly dT and Superscript RNaseH- reverse transcriptase (Gibco-BRL, USA). Subtractive hybridization was performed between biotin labeled driver sscDNA (unstressed) and tester (stressed) mRNA at 65oC for 1 hour. The tester-driver hybrids and excess of the driver were separated from the unhybridized tester mRNA using streptavidin paramagnetic particles. Tester mRNAs that did not hybridize with the driver were used for cDNA synthesis (Stratagene). A primary cDNA library was produced using a lambda ZAP XR cDNA synthesis kit (Stratagene). Randomly selected clones amplified using M13F and M13R-primers, PCR product was purified using PCR purification system (Stratagene) and sequenced from the 3' end on an MegaBACE 1000 DNA analysis system (Amersham Pharmacia, Uppsala, Sweden). Sequences are processed using the VecScreen (http://www.ncbi.nlm.nih.gov/VecScreen.html) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained at least 100 bases were submitted to GenBank.
characteristics.characteristic: description
characteristics.text: 21 day-old seedlings
characteristics.characteristic: dev-stage
characteristics.text: E.coli
characteristics.characteristic: lab-host
characteristics.text: Carthamus tinctorius
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_4222
characteristics.characteristic: organism
characteristics.text: EcoRI
characteristics.characteristic: re_1
characteristics.text: XhoI
characteristics.characteristic: re_2
characteristics.text: Leaf
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/PO_0025034
characteristics.characteristic: tissue-type
characteristics.text: Safflower (A-1) drought stressed subtracted cDNA library
characteristics.characteristic: title
characteristics.text: Lambda Uni-ZAP XR
characteristics.characteristic: vector