LIBEST_026414 Chickpea drought stressed cDNA SSH library BULK1

Overview
Library NameLIBEST_026414 Chickpea drought stressed cDNA SSH library BULK1
Unique NameLIBEST_026414
OrganismCicer arietinum (chickpea)
Typecdna_library
LIBEST_026414 Chickpea drought stressed cDNA SSH library BULK1
  • Cultivar: RILs derived from cross between ICC4958 and ICC1882
  • Tissue: Root
  • Developmental stage: Flowering stage
  • Vector: pGEM-T Easy vector
  • Detail: A RIL population was derived from cross between ICC4958 a high root biomass genotype (HRB) and ICC1882 a low root biomass(LRB) genotype. 10 extreme RILs exhibiting HRB and LRB were selected. Bulks of the 10 RILs were used for construction of cDNA SSH library. RILs were grown in pots under near normal optimal condition in glasshouse. Control plants (well watered) were maintained close to 80% field capacity by keeping the pot wet to that level every day by compensating for the loss due to transpiration and water stressed plants were exposed to gradual water stress by partly compensated for the water loss due to transpiration. Plant samples from the control (well watered) and stressed plants were harvested when the transpiration ratio reached to 0.1 in the water stressed plants. Total RNA from root tissue was isolated using Trizol reagent and equal amount of total RNA from 10 HRB RIL and 10 LRB were mixed to produce two Bulks of RNA, further mRNA was extracted from the total RNA using poly(A) tract mRNA Purification Kit (Promega). Forward SSH library was constructed using subtraction of cDNA synthesized from water stressed bulk of 10 HRB RILs plants as tester and that from the bulk of 10 RILs with LRB plants as driver with the Clontech PCR-Select cDNA subtraction Kit (CLONTECH, USA) according to the manufacturer‘s protocol. Differentially expressed cDNAs were cloned in pGEMT-Easy vector and transformed into E.Coli. Sanger sequencing of the selected clones were done using universal sequencing primers T7, SP6 and M13.
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