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Expression levels of these 16 genes, determined by Taq Man® RT-PCR analyses and Affymetrix Gene Chip® arrays, were normalized and tested for overall variance and equivalence of the means.Pair-wise analysis showed that two reference genes were sufficient for proper normalization, except when analyzing the gene expression studies in all samples set.Results of this study can help in the selection of reference genes for quantitative real time PCR (q RT-PCR) normalization in pearl millet that will contribute towards more accurate and reliable quantification of transcripts in this important crop of the drylands.To compensate for potential variability introduced by the procedure, normalization of target gene expression has been established.Accurate normalization has become an absolute prerequisite for the correct quantification of gene expression. cookies, tags, and tracking settings to store information that help give you the very best browsing experience.To understand more about cookies, tags, and tracking, see our Privacy Statement *Prices in US$ apply to orders placed in the Americas only.Data obtained in the quantitation, after its normalization, provide a powerful source of gene expresion patterns [10].Different housekeeping genes are used as internal controls for RT-q PCR analysis [11-13].The results and parameters of expression profile data are comparable to the two-step real-time PCR, that were previously reported for , L; HMG Co A reductase; Pollen allergens; Expression; RT-PCR HMG Co A Reductase: 3-Hydroxy-3-Methylglutaryl Coenzyme A Reductase; m RNA: messenger Ribonucleic Acid; NCBI: National Centre of Biotechnology Information; RT-PCR: Reverse Transcribed Polymerase Chain Reaction; RT-q PCR: Real-Time quantitative Polymerase Chain Reaction Respiratory diseases with a background in pollen allergens possess an increased tendency throughout the urbanized area.Pollen allergic diseases are the result of hypersensitivity to aeroallergens and are reported as to affecting up to the 30% of population [1,2]. Both of them were reported as to be related to the birch pollen allergens Bet V1 and Bet V2 [7]. Thinking of above mentioned circumstances, developing of reproducible and reliable methods of the allergenic potencial of different sources of spring pollinosis will lead to their better management.Several strategies are in use for the normalization of data, including normalization to sample size, to total RNA or to an internal reference.Among these, the use of housekeeping genes as an internal (endogenous) control is the most common approach.However, the accuracy of this technique depends on the quality of internal controls.Endogenous internal controls, often referred to as ‘housekeeping’ or ‘reference’ genes, offer multiple practical advantages and are therefore widely used in research and clinical applications.

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