Quality testing & analysis of bt-cotton hybrid seeds

Abstract

This Internship focuses on the detection of the Cry gene in Bt cotton seeds using Sandwich ELISA and the evaluation of seed germination using cellulose paper. The main purpose is to determine whether the Bt cotton seed contains the Cry gene, which encodes the production of an antibiotic that protects the plant from pests, and to evaluate the potency of the seed using germination techniques. This method includes the preparation of seed extracts for sandwich ELISA and germination testing using cellulose paper. The extracted proteins are subjected to sandwich ELISA, which involves the use of specific antibodies to detect the presence of Cry proteins in the Sample. Germination tests were performed using water-moistened cellulose paper and were used to measure seed viability based on seedling emergence. In the ELISA sandwich test, samples are prepared by grinding the seeds into a fine powder and extracting the proteins in a buffered solution. Protein concentrations were then determined using the Bradford assay. The sample is incubated with an antibody specific to the Cry protein followed by a second antibody conjugated to the enzyme. When substrate is added, the enzyme catalyzes a colorimetric reaction, allowing Cry proteins to be detected in the sample. The assay was optimized for sensitivity and specificity, and the results were analyzed to determine the presence of the Cry gene in Bt cotton seeds. In the Germination test, cellulose paper is placed in a Petri dish with a layer of filter paper on top. The filter paper was moistened with water and the seeds were evenly spread on the filter paper. The Petri dish is covered with a plastic sheet to retain moisture and the seeds are allowed to germinate for 7 days. The occurrence of genes is observed and calculated for the evaluation of gene strength. The internship led to several major discoveries, including the successful detection of Cry proteins in Bt cotton seeds using a sandwich ELISA. The Results showed that the seeds had different Cry protein levels depending on the variety of Bt cotton. In addition, germination tests showed increased growth of Bt cotton seeds, indicating their full viability. These findings have important implications for the development of better farming techniques and for ensuring the quality of cotton seeds. Confirming the presence and potential of the Cry gene in Bt cotton seeds by germination tests, this study adds to current knowledge about the properties and potential of Bt cotton as a crop.