|Library Name||LIBEST_026409 Chickpea drought stressed cDNA SSH library AB2 |
|Unique Name||LIBEST_026409 |
|Organism||Cicer arietinum (chickpea) |
|Type||cdna_library |LIBEST_026409 Chickpea drought stressed cDNA SSH library AB2
- Cultivar: ICC4958 and ICC1882
- Tissue: Root
- Developmental stage: Flowering stage
- Vector: pGEM-T Easy vector
- Detail: Chickpea drought tolerant cultivar ICC4958 and susceptible cultivar ICCC1882 was used in this study. Plants 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 mRNA was extracted from the total RNA using poly(A) tract mRNA Purification Kit (Promega). Reverse SSH library was constructed using subtraction of cDNA synthesized from water stressed ICC1882 plants as tester and that from the water stressed ICC4958 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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