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Articles by K. Shahrokhabadi
Total Records ( 2 ) for K. Shahrokhabadi
  K. Shahrokhabadi , R.T. Afshari , H. Alizade , J.T. Afshari and G.R. Javadi
  This report described an improved method for isolating intact purified RNA from freezing organs of bread wheat plants. High-quality RNA is important in studying gene expression. Common RNA extraction protocols have produced poor yields because freezing leaves contain polysaccharides and RNases. We used two methods for isolating RNA and comprised them. CTAB (cetyltrimethylammonium bromide) method protocol is based on a guanidine thiocyanate extraction combined with additional purification steps using butanol and the ionic detergent CTAB. Using this protocol, RNA yields ranged from 40-70 μg of total RNA 200 mg of fresh tissue. This method can be adapted to large-scale isolations, allowing the recovery of larger amounts of intact RNA (up to 250 μg g-1 of fresh tissue).
  K. Shahrokhabadi , R.T. Afshari , H. Alizade , J.T. Afshari and G.R. Javadi
  In this study, we exploit the useful described CODEHOP primer design and RT-PCR strategy for targeted isolation of homologues in large gene families. The method was tested with two different objectives. The first was to apply CODEHOP strategy for design degenerate oligonucleotide primers in a broad range of plant species. The second was to isolate an orthologus of the transcription factor of dehydration-responsive element binding protein (DREB) and to determine the complexity of gene family in bread wheat. We used a new primer design strategy for PCR amplification of unknown targets that are related to multiply-aligned protein sequences. Each primer consists of a short 3` degenerate core region and a longer 5` consensus clamp region. Only 3-4 highly conserved amino acid residues are necessary for design of the core, which is stabilized by the clamp annealing to templates molecules. This provides the possibility of isolating numerous additional DREB genes by Polymerase Chain Reaction (PCR) with degenerate oligonucleotide primers. The relationship of the amplified products to DREB genes was evaluated by several sequence and genetic criteria. Present data show that expression of DREB and its homologues, is induced by low temperature stress. Towards this step, it found that the expression of DRE-regulated genes increased freezing tolerance in plants.
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