DETECTION OF RICE SPS GENE BY SIMPLEX PCR METHOD | ||
| IRAQ JOURNAL OF AGRICULTURE | ||
| Article 1, Volume 19, Issue 6, October 2014, Pages 0-0 | ||
| Authors | ||
| A. N. Thwainy; Y. I. Farhan; S. S. Fakhry | ||
| Abstract | ||
| The study include 14 samples were chosen for sucrose phosphate synthase - sps gene. Molecular characterization which were characterized by PCR using SPS primer to identify up to generic level. Genomic DNA purification of 14 parallel Rice samples was carried out by DNeasy Plant Kit. The Yields and purities of the purified genomic DNAs were measured by spectrophotometer. The purities of all samples were higher than 1.7, which clearly demonstrates that the purified genomic DNA is of high quality. The integrity of the purified genomic DNA was also analyzed by agarose gel electrophoresis. The genomic DNA was used as template for PCR utilizing primers specific for the sps gene, and the amplified PCR products size were about 251base pair and the results demonstrate that the purity of the purified genomic DNA is sufficiently high for a sensitive PCR analysis. Detection primer with the optimized quantity of 10 pmole, were used in the PCR. PCR products treated with that primer pairs were observed only in rice samples. The present study confirmed that detection primer pair could efficiently amplify the corresponding gene. Nonspecific bands were not present at the annealing temperature 60 °C and the sps internal gene was amplified in all the rice samples with a size of 251 bp. For rice specific genes. PCR products treated with SPS primer pairs were observed. We showed that the PCR detection system used in the present study under optimized condition could be an appropriate tool, without primer interference or dimerization, for the detection of GM rice events. Here, we present results that will be useful for GM crop assessment in detecting multiple targets simultaneously with simplex PCR-based methods for various purposes, such as a part of the approval process or post-market monitoring. | ||
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