Medical Biochemistry for Embryo and Starchy - MyAssignmenthelp.com

Question: Discuss about theMedical Biochemistry for Embryo and Starchy. Answer: Introduction A wheat kernel has three main sections- embryo, starchy endosperm and protective layer. For a plant to carry out germination, many cells and cellular materials reproduce from the original embryo. Acid phosphate is a major enzyme produced for the initial stage of germination. This enzyme is responsible for breaking down phosphate esters and releasing the phosphate for necessary metabolic activities. In this experiment acid phosphate is to be extracted from wheat germ and its enzymatic activity is to be determined. The results would be then compared with the standard graphs produced in experiment 3A and the amount of acid phosphate would be determined. Materials Chemicals/ biological ingredients Hazard safety check Wheat germs Raw wheat germ is to be stored sealed in a vaccum storage container to avoid heat, humidity and exposure to air 50 mM Sodium Acetate Buffer (NaOAc) solution Protective clothing and gloves are to be worn while handing it. Avoid breathing it and store away from moisture p-nitrophenol phosphate (PNPP) stock solution (1 mg/mL) Handle in accordance with good industrial hygiene and safety practice. Personal protective equipment might be used Deionised H2O (dH2O) Good hygiene procedures are to be followed and splashing and spraying are to be avoided 1M NaOH Container to be kept dry and water is not to be added to the solution. Avoid contact with skin (Keith, 2016) Instruments Conical centrifugation tube Disposal of used tubes is necessary for avoiding contamination from used subtances Microcentrifuge tube Avoid spillage of used substances. Disposal of used tubes is necessary for avoiding contamination High velocity centrifuge (max speed of 13,000 rpm) After centrifugation is carried out it is necessary to take the rotor to a biosafety cabinet prior to removal of the lids. If there is a leak in the centrifuge appropriate steps are to be taken Microplate reader Repeated exposure to be avoided and protective equipments to be used. Container to be placed in dry, well-ventilated place Benchtop vortex Avoid contact with surface, ground water or soild. Contact wtith skin is to be avoided and adequate ventilation is to be maintained Materials Chemicals/ biological ingredients Hazard safety check Bradford reagent Store in tightly closed container at 2-8C. Bovine serum albumin (BSA) stock solution (1mg/mL) To be used in well ventilated areas and personal protective equipment to be worn Instruments 1 mL Cuvettes To be used dry and spillage to be avoided Spectrophotometer All operations are to be performed uing gloves and safety goggles. The instrument is to be used in a clean environment away from other instruments that cause vibration. Mechanical compoenents are to be maintained in good condition (Scopes, 2013). Introduction It is crucial to determine the molecular and physical properties of a protein in biochemistry for understanding the unique characteristics and all possible downstream application. Electrophoresis is a commonly used method in biochemistry for separating protein and nucleic acid. One significant method for purifying proteins is using polyacrylamide gel electrophoresis (PAGE). The widely used method, called protein denaturation, applies sodium dodecyl sulphate for stripping down the protein into linear amino acid sequence for producing better migration pattern in the gel. This is termed as SDS-PAGE process. From previous In this experiment the proteins within the extract would be separated through SDS-PAGE method for determining the different forms of protein present within the wheat germ extract. Materials Chemicals/ biological ingredients Hazard safety check 10% Resolving gel Proper personal protective equipment to be used while handling the agent Stacking gel Proper personal protective equipment to be used while handling the agent Stock loading buffer solution (5x and 2x) Proper personal protective equipment to be used while handling the agent Electrophoresis tank buffer Proper personal protective equipment to be used while handling the agent (Keith et al., 2005) Coomassie Staining Solution Proper personal protective equipment to be used while handling the agent for avoiding unnecessary staining Pure acid phosphatase enzyme (1 mg/mL) Repeated exposure to be avoided and spills to be avoided Wheat germs Raw wheat germ is to be stored sealed in a vaccum storage container to avoid heat, humidity and exposure to air 50mM Sodium Acetate Buffer (NaOAc) stock solution Protective clothing and gloves are to be worn while handing it. Avoid breathing it and store away from moisture (Hegyi et al., 2013) Instruments Electrophoresis apparatus Equipment and bench tops to be decontaminated using soap and water. Disposal of all contaminated disposables necessary Gel loading tips After loading of gel the tips are to be discarded away for avoiding contamination Weight boats To be kept away from high heat 100 C heating block After usage of the block rapid and sufficient collign of the block is necessary High velocity centrifuge (max speed of 13,000 rpm) After centrifugation is carried out it is necessary to take the rotor to a biosafety cabinet prior to removal of the lids. If there is a leak in the centrifuge appropriate steps are to be taken Microcentrifuge tubes Avoid spillage of used substances. Disposal of used tubes is necessary for avoiding contamination (Chawla, 2014) References Chawla, R. (2014).Practical clinical biochemistry: methods and interpretations. JP Medical Ltd. Hegyi, G., Kardos, J., Kovcs, M., Mlnsi-Csizmadia, A., Nyitray, L., Pl, G., ... Venekei, I. (2013). Introduction to Practical Biochemistry.ELTE Faculty of Natural Sciences, Institute of Biology. Keith (Ed.) Wilson, John (Ed.) Walker. (2005).Practical Biochemistry: Principles And Techniques. Cambridge University Press. Keith, W. (2017). A biologist's guide to principles and techniques of practical biochemistry. Scopes, R. K. (2013).Protein purification: principles and practice. Springer Science Business Media.