T1 lines (G1‚ G2‚ G3‚ G4 and G5) maintained in greenhouse were used for extraction of soluble protein. Lectin was extracted from leaves (2g) in 2 ml of TBS containing 0.25mM isopropylthio-ß-D-galactoside (IPTG; Merck) for overnight at 4 °C‚ the homogenate was centrifuged at 6700g for 20 min at 4° C. The supernatant collected was used for hemagglutination assay. The concentration of total soluble protein was determined by the Bradford method (1976).Further the quantity of recSRL expressed in transgenic
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Background on Genomic DNA Isolation and Purification Generally‚ all methods involve the disruption and lysis of cells. This is followed sometimes by the removal of RNA (by RNAses‚ salt or other methods). Choosing which method to use will depend on many selection factors including: DNA is isolated from proteins by several methods including digestion of proteins by the enzyme proteinase K. Proteins are removed subsequently by salting-out‚ organic extraction‚ or binding of the DNA to a solid-phase
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Concentration‚ pH and Temperature on Enzyme Activity” Abstract: In the following experiments we will measure precise amounts of potato extract as well as Phenylthiourea‚ combined with or without deionized water and in some instances change the temperature and observe and record the reaction. We will also investigate the different levels of prepared pH on varying samples of the potato extract and the Phenylthiourea and record the results. We will answer question such as what is the best temperature for optimum
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Effects of Temperature‚ pH‚ Enzyme Concentration‚ and Substrate Concentration on Enzymatic Activity INTRODUCTION Enzymes‚ proteins that act as catalysts‚ are the most important type of protein[1]. Catalysts speed up chemical reactions and can go without being used up or changed [3] Without enzymes‚ the biochemical reactions that take place will react too slowly to keep up with the metabolic needs and the life functions of organisms. Catecholase is a reaction between oxygen and catechol
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was inserted into the spectrophotometer and measured with the 420 nm wavelength. This procedure was repeated for the remaining test tubes. Exercise B required 4 test tubes labeled with “A‚B‚C‚ or D” and they held the solution of different pH buffer and potato juice volumes‚ and 1 mL of water (Table 4). These tubes were the blanks for the spectrophotometer for recalibrating it. The additional 4 experimental test tubes were composed of the same contents but 1 mL of the substrate catechol was added instead
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5. RESULT AND DISCUSSION RESULT The enzyme 6-phosphogluconate is the significant enzyme in the oxidation phase of pentose phosphate pathway. The homogenate was made by adding the buffer to the liver of the quail after one hour centrifuge at 12300 RPM materializing at 4°C. Precipitation protein was working by ammonium sulfate; it loaded directly to column 2’‚ 5’ ADP Sepharose 4B affinity chromatography‚ which has been a high affinity to NADP+ substrate. The flow rate of elution was 23 ml/hr
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phosphate buffer. 10 drops of catechol and 10 drops of potato juice‚ which contains catecholase‚ were added to each of the seven tubes. The tubes were covered with Parafilm‚ stood for 5 minutes‚ and mixed every minute. After the 5 minutes‚ data was recorded based on the intensity of color with the “0‚+‚++‚+++”
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substance that can be formed during aerobic respiration and catalase removes this product. The activity of catalase can be measured by finding the rate of oxygen release from hydrogen peroxide. Potato provides a suitable source of catalase and the pH is varied in this experiment using citric acid-sodium phosphate buffer solutions at pH values of 4.4‚ 5.2‚ 6.5 and 7.5. Catalase is an enzyme‚ a biological (organic) catalyst. Hydrogen peroxide is the substrate for catalase. It lowers the activation energy required
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reactant. The enzyme which will be used is different concentrations of potato and the reactant used will be Hydrogen Peroxide. Hydrogen Peroxide which will be the buffer solution is a PH of 7.2. My hypothesis for the experiment is that as the concentration of the enzyme is increased the rate of reaction will be increased‚ producing oxygen at a faster rate. The results in my opinion will show positive correlation. The amount of buffer solution (hydrogen peroxide) will be kept the same throughout the experiment
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disrupting the membranes with a solution of detergent and salt‚creating a cell homogenate. Once the DNA is released from the nucleus‚ it must be protected from nucleases‚ enzymes which will degrade the DNA. Keeping the cell homogenate cold and various chemical components of the homogenization medium help restrict the action of these nucleases. The final step of this protocol involves precipitation of DNA from the homogenate. When the homogenization medium is added in the first step‚ the positive ions
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