Explain the source of errorsources of error- in relation to the assay, errors can be from extraction method or the assay itself (like use of spectro), explain the effect of the error especially to the absorbance SFor animal tissues-Homogenize 2 grams of the tissue using a mortar and pestle.Get one gram of the tissue sample and add 5 mL of 7.5% trichloroacetic acidsolution or 5mL of 20% glacial acetic acid solution. Cover the tube with parafilm.Agitate the mixture for 10 minutes using a vortex.Filter the mixture using Whatman 1 filter paper.Centrifuge the sample at 4000 rpm for 3 minutes.Separate the supernatant and discard the pellet.If there is substantial turbidity, increase the settings to 5000rpm and centrifuge foranother 3 minutes.Collect the supernatant and place it into another test tube.Label the tube(s) accordingly.For liquid samplesIn a test tube, mix 1 mL of oil sample with 5mL of distilled water. Cover the tubewith parafilm.Agitate the mixture for 10 minutes using a vortex.Allow the mixture to settle, until the two phases separate.Collect the aqueous layer and place it into another test tube and cover it withparafilm.Label the tube(s) accordingly.NOTE: If the samples cannot be analyzed immediately, add 500μL of 0.5% BHT or 5.0% gallic acidsolution to prevent further oxidation. Refrigerate the samples at 4°C prior to analysis the next day.C. TBARS Assay1. In a test tube, mix 1 mL of the samples and 1 mL of thiobarbituric acid reagent.2. Place the test tube in a water bath (95°C) for 60 minutes to develop a color reaction. Maintainthis temperature throughout the procedure.3. Cool the sample.4. Measure the absorbance at 532nm. Blank the sample using the absorbance of the samplewithout the thiobarbituric acid solution. Follow the procedure for calibrating the cuvettes andthe spectrophotometer.5. Determine the concentration based on the obtained linear regression equation.6. Perform three replicates per sample. Express the concentrations as mean ± standard deviationof the unit of concentration, based on how you prepared the calibration curve.Then, express the final concentration as mM MDA equivalents per gram of the sample. Performa T-test to your data.NOTE: If the concentration of the sample is too high, dilute the sampie and perform theprocedure again.In your laboratory report, focus on the following concepts:a. Principle of the TBARS assayb.Sources of errorc. Comparison of results based on the type of sampled. Efficiency of extraction methode. Limitations of the assayf. Applications to health decisions (based on results)A

Sources of error can be defined as the factors which affect the accuracy and precision of an assay.…

C. TBARS Assay 1. In a test tube, mix 1 mL of the samples and 1 mL of thiobarbituric acid reagent. 2. Place the test tube in a water bath (95°C) for 60 minutes to develop a color reaction. Maintain this temperature throughout the procedure. 3. Cocl the sample. 4. Measure the absorbance at 532nm. Blank the sample using the absorbance of the sample without the thiobarbituric acid solution. Follow the procedure for calibrating the cuveties and the spectrophotometer. 5. Determine the concentration based on the obtained linear regression equation. 6. Perform three replicates per sample. Express the concentrations as mean ± standard deviation of the unit of concentration, based on how you prepared the calibration curve. Then, express the final concentration as mM MDA equivalents per gram of the sample. Perform a T-test to your data. NOTE: If the concentration of the sample is too high, dilute the sampie and perform the procedure again. In your laboratory report, focus on the foilowing concepts: a. Principle of the TBARS assay b. Sources of error c. d. e. Limitations of the assay f. Applications to health decisions (based on results) Comparison of results based on the type of sample Efficiency of extraction method

C. TBARS Assay 1. In a test tube, mix 1 mL of the samples and 1 mL of thiobarbituric acid reagent. 2. Place the test tube in a water bath (95°C) for 60 minutes to develop a color reaction. Maintain this temperature throughout the procedure. 3. Cocl the sample. 4. Measure the absorbance at 532nm. Blank the sample using the absorbance of the sample without the thiobarbituric acid solution. Follow the procedure for calibrating the cuveties and the spectrophotometer. 5. Determine the concentration based on the obtained linear regression equation. 6. Perform three replicates per sample. Express the concentrations as mean ± standard deviation of the unit of concentration, based on how you prepared the calibration curve. Then, express the final concentration as mM MDA equivalents per gram of the sample. Perform a T-test to your data. NOTE: If the concentration of the sample is too high, dilute the sampie and perform the procedure again. In your laboratory report, focus on the foilowing concepts: a. Principle of the TBARS assay b. Sources of error c. d. e. Limitations of the assay f. Applications to health decisions (based on results) Comparison of results based on the type of sample Efficiency of extraction method

Biochemistry

9th Edition

ISBN:9781319114671

Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.

Chapter1: Biochemistry: An Evolving Science

Section: Chapter Questions

Problem 1P

See similar textbooks

Similar questions

Helping tags: Biology, Food Microbiology, Spore formers, sulfide spoilage spores 25 g of sugar sample was obtained from the production line for analysis of raw ingredients. It was diluted to 250 ml and was heated at 100 deg Celsius for 5 minutes. After cooling, 20 ml of the sample solution was divided equally among 6 tubes of sulfite agar. After incubation, the following counts were obtained: 35, 26, 29, 32, 34, and 31. Compute for the number of sulfide spoilage spores and express the count per 50 g of sugar. . . . WILL UPVOTE, just pls help me answer the question and show COMPLETE solution.
Procedure Explant Preparation and Inoculation 1) The leaf explants were cut into squares (approximate size = 1cm x 1cm ). 2) The explants were soaked in 95% EtOH for 3 minutes with constant shaking. 3) EtOH was then decanted and 10% of Chlorox bleach was added together with a few drops of Tween 20, with constant shaking. 4) The explants were rinsed with autoclaved distilled water for at least 5 times. 5) The explants were then left to dry in the laminar air flow cabinet. 6) The explants were cultured in the appropriate media (2 explants in basal MS without BAP and the other two in MS with BAP). 7) Three explants were placed on its upper surface down and the other three with lower surface down in both basal MS media with and without BAP. 8) All the culture jars (name, date and treatments employed etc.)were labelled and incubated them (in dark) in the growth chamber or culture room. 9) Explants were placed on the surface of the medium and the Petri dish were sealed with parafilm. 10)…
EXPERIMENT:EFFECT OF CARBON SOURCES ON THE MICROBIAL GROWTH. 1.OBJECTIVE To study the effect of media components on microbial growth 2.PROCEDURES Preparation of fungal suspension inoculums Prepare the laminar cabinet for the inoculum preparation by applying normal aseptic techniques To a full-grown fungal Petri dish, add 25ml sterilized distilled water and rub the agar gently with glass rod Pour the suspension into an empty flask and continue previous steps to achieve your desired volume of inoculums suspension. Cotton-plugged and aluminium-wrapped the mouth of inoculum flask. For certain fungal species like Niger, the suspension from the agar should be filtered upon pouring into the inoculum’s flask. Preparation of salt media of different carbon sources. Prepare the Modified Czapek Dox media by weighing the following salts according to your desired working volume. KH2PO4 (1 g/L) NaNO3 (3 g/L) MgSO4.7H2O (0.5 g/L) Carbon source (30 g/L) KCl (0.5 g/L) 2.…
True or false ? images will appear brighter when light waves are in phase. the condenser portion of the microacope focuses light on the speciment.
A microbiologist has isolated a new strain of bacteria, and is trying to characterize aspects of its growth. Three flasks of the same media have different amounts of NaCl added. Other growth conditions are identical, and the flasks inoculated at the same time. At regular intervals over a 3 hour period, culture samples are taken and turbidity is measured. Here are the absorbance values over time in a table and graphed. Absorbance (OD600) Time (min) 20% NaCl 3% NaCl 0% NaCl 0 0.05 0.05 0.05 30 0.06 0.05 0.05 60 0.075 0.06 0.05 90 0.11 0.07 0.049 120 0.25 0.1 0.048 150 0.5 0.105 0.055 180 0.7 0.11 0.055 210 0.99 0.15 0.06 240 1.22 0.18 0.055 270 1.35 0.19 0.06 300 1.5 0.2 0.06 330 1.6 0.25 0.062 360 1.7 0.3 0.06 Based on these data, which of the following can be determined about this bacterium? It is psychrophilic. It…
Pattern used to inoculate agar slant to get luxuriant culture. This pattern increases the surface area of agar that can be inoculated.
The following figures are the photomicrographs of the samples taken with Kern&Sohn Camera Microscope at 10x magnification. Samples were stained with Lugols solution for better viewing. Differentiate these two image
ITRATE TESTThis test determines the ability of bacteria to use citrate as source of carbon andinorganic ammonium salt as a source of nitrogen. The following are the stepwise procedurein conducting citrate test:1. Infuse the Simmons citrate agar on the slant by touching the tip of sterilized needleto a colony.2. Incubate at 37 degree Celsius for 24 to 48 hours.A positive result is indicated by the pH indicator, bromothymol blue which turns the growthgreen to blue. Task: In a tabulated form, determine if Salmonella typhi the are positive or negative withcitrate test. Interpret your answer and include the reaction of organism on this test.
Kligler’s iron agar and SIM are multiple test media.. What component of both media allows you to detect the preceding characteristic?
Heat-labile material to be introduced to culture medium was filtered using membrane filters with unknown pore size. Effect: Rationale:
Create a flow chart to perform the required tests to identify unknown microorganism: - Use at least 3 other biochemical tests from the following list: Mannitol salt agar, Blood agar, Starch agar, Tributyrin Agar, Gelatin, Casein agar, Indole Production, MR-VP, Citrate, Hydrogen Sulfide test, Urease test, Nitrate reduction test, Catalase test, and Oxidase test. The Gram positive organisms that could be present in your unknown mixture are: Bacillus cereus, Micrococcus luteus, Staphylococcus aureus, Staphylococcus epidermidis and Lactococcus lactis.
Multiple Matching. For each type of medium, select all descriptions that fit. For media that fit more than one description, briefly explain why this is the case. _______mannitol salt agar a. selective medium _______chocolate agar b. differential medium _______MacConkey agar c. chemically defined (synthetic) _______nutrient broth medium _______brain-heart infusion broth d. enriched medium _______Sabouraud’s agar e. general-purpose medium ________triple-sugar iron agar f. complex medium ________SIM medium g. transport medium