characteristics.text: LIBEST_017692
characteristics.characteristic: External Id
characteristics.text: SAMN00149812
characteristics.characteristic: External Id
characteristics.text: 2014-02-03T23:44:04.367Z
characteristics.characteristic: INSDC first public
characteristics.text: 2015-05-20T14:15:14.660Z
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: VZM-1
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 allowed to grow for fourteen days. The pots were maintained in the departmental botanic garden under natural photoperiod (10-12 hrs, 28 ( 4oC). Seedlings were maintained at 100% field capacity by watering daily. Fourteen day-old plants were subjected to moisture stress by withholding water. Leaf samples collected from unstressed (100% FC) and stressed (once field capacity reached 25%) plants, flash frozen and stored at -80oC. The frozen samples were used for RNA extraction (LiCl2 method). PolyA mRNA was isolated using streptavidin paramagnetic particles (SPMPs; 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 ss cDNA(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 (SPMPs; Roche, Germany). 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 partially sequenced from the 3' end on an MegaBACE 1000 DNA analysis system (Amersham Pharmacia, Uppsala, Sweden). Sequences are processed using the HarvEST pipeline (http://harvest.ucr.edu) 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 deposited to GenBank. Sequences that retained at least 100 bases were deposited to GenBank.
characteristics.characteristic: description
characteristics.text: 24 days seedlings
characteristics.characteristic: dev-stage
characteristics.text: Ecoli
characteristics.characteristic: lab-host
characteristics.text: Macrotyloma uniflorum
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_271171
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: Horsegram (VZM-1) drought stressed subtracted cDNA 24 days seedlings
characteristics.characteristic: title
characteristics.text: Phagemid pBluescript SK(-)
characteristics.characteristic: v_type
characteristics.text: Lambda Uni-ZAP XR
characteristics.characteristic: vector