Week 5 LabOL_Pipetting

bio 2 from Lab Manual 4 TH edition Leboffe and Pierce

General Electrophoresis Questions: 1. What makes macromolecules move through the gel in electrophoresis?2. What determines the speed at which macromolecules move through the gel in electrophoresis? In a single gel, why do some move faster than others?3. Why do we use different procedures for DNA and protein electrophoresis?

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Helping tags: Biology, bacteria, lag phase WILL UPVOTE, just pls help me answer the following questions and explain them clearly. Thank you. TOPIC: Bacterial Growth Curve 1. How may the following growth and culture conditions affect the length of the lag phase? EXPLAIN YOUR ANSWERS. a) inoculum is from an old culture b) shifting cells from rich culture medium to a poorer one c) exponentially growing culture is transferred into the same medium under the same growth conditions

Discuss the principles and applications of Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry in molecular diagnostics.

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Topic: Isolation of Crude Ovalbumin from Egg White by Ammonium Sulfate Precipitation(Salting Out) Egg whites contain about 88.5% water and about 10% proteins, from which ovalbumin is the major component. The eggs were cracked open, and the egg whites were separated carefully from the yolk. Stirring of the egg whites was done to break up the membrane. This was followed by filtration through cheesecloth, where a clear filtrate was obtained. About 40.0 mL of the filtered undiluted egg white was measured, and 0.10 mL of 1 M acetic acid was slowly added for every 1.0 mL of egg white with constant stirring. A turbid mixture with few amounts of white precipitate may be expected to form. These may be removed by filtering the mixture using a cheesecloth After filtration, 30.0 mL of the filtrate was obtained, and the solution was brought from 0% to 40% saturation by adding the required amount of powdered ammonium sulfate, (NH4)2SO4 After adding ammonium sulfate, the mixture was allowed to stand…

Q1: Why are E.coli cells subjected to heat shock induction when the optical density of the bacterial culture has reached 0.6-0.8? Q2: Why are these wash steps critical for the preparation of protein samples used in 2D gel electrophoresis?

biomedical instrumentation question

Assignment_12.pdf - Adobe Reader File Edit View Window Help Оpen 1 / 1 99,1% Tools Fill & Sign Comment 3. Recombinant protein is produced by a genetically engineered strain of Escherichia coli during cell growth. The recombinant protein can be considered a product of cell culture even though it is not secreted from the cells; it is synthesized in addition to normal E. coli biomass. Ammonia is used as the nitrogen source for aerobic respiration of glucose. The recombinant protein has an overall formula of CH1.5500.31N..25. The yield of biomass (excluding recombinant protein) from glucose is measured as o.48 g/g; the yield of recombinant protein from glucose is about 20% of that for cells. How much ammonia is required? What is the oxygen demand? If the biomass yield remains at 0.48 g /g, how much different are the ammonia and oxygen requirements for wild-type E. coli that is unable to synthesize recombinant protein? а. b. с. 24°C 08:39 Berawan 27/04/2022

NEED ASAP Using the list provided (note you may not need all the steps listed) state the steps in cDNA production 1. Confirm size of bands by comparing to molecular size marker. 2. Ligate oligonucleotides onto blunt ended product. 3. Cut the cDNA with a unique restriction enzyme. 4. Ligate the cDNA and vector followed by transformation of E. coli. 5. Plate cells onto LB-ampicillin plates containing X-gal. 6. Choose colonies that are white in colour. 7. Cut plasmid with unique restriction enzymes. 8. The size of the clone is small and it will ensure that the entire gene will be found in one fragment. 9. Employ replica plating technique and select cells that are ampicillin-sensitive and tetracycline resistant. 10. Blot petri plates with a nitrocellulose filter, lyse the cells and denature the DNA with a dilute sodium hydroxide solution. 11. Blot electrophoresis gel with nitrocellulose filter and denature the DNA with a dilute sodium hydroxide solution. 12. Flood the filters with…

Q24:  A 7 dilution is performed on a culture of bacteria in order to perform viable plate counts. From the dilution, *0.1 mL* of solution is plated on solid media, and 31 colonies of bacteria grow on the petri dish.  Assuming each colony came from a single bacterium, how many bacteria are in a single mL of the original culture? Express your answer to two decimal places using standard notation. Since only 0.1 mL is put on the plate, this counts as an extra dilution!!! Any time less than 1 mL is transfered, a dilution is being performed. Any time more than 1 mL is transfered, a concentration is being performed.

Enhancement Questions:  What do you think are the crucial steps in Polymerase Chain Reaction technique? Enumerate the advantage and disadvantage of Polymerase Chain Reaction technique?

Q21:  Polymerase Chain Reaction (PCR) (Links to an external site.) is an essential technique for the study of DNA. PCR uses a DNA polymerase enzyme from Thermophilus aquaticus (Links to an external site.) to amplify DNA extracted from any sample. In order for the reaction to work, researchers must transfer microliters of sample to the reaction vessel for PCR. If a researcher has a DNA sample with a concentration of 0.2 micrograms per microliter, how many microliters of DNA must be transfered to conduct a 50 microliter reaction if the final concentration should be 2.3 nanograms per microliter? Report your answer to two decimal places using standard decimal notation (e.g. 0.5, not 5.0e-1)

topic: sds-page gel If APS is not available, what other chemicals can be used alternatively to initiate the polymerization?

please answer for all three questions  asap  all three questions are related

present briefly the principle of size exclusion chromatography.  give a brief description of the use of protein assays.

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Topic: Isolation of E. coli bacteriophage   What would happen if: - the enrichment method in the isolation of bacteriophage was omitted? - the chloroform was not added to the enrichment? - the 0.1 ml lysate-E. coli mix was plated directly on top of the bottom agar?

topic: gel electrophoresis What is the purpose of the running buffer?

Correctly identify the steps and outcomes in the process of screening for a nutritional mutant of a bacterium. Drag the appropriate labels to their respective targets.

Discuss shortly five important things that should be kept in mind when designing a centrifugation protocol?

Question questions. : Answer the following A) Compare between the two major types of cells. Also, explain the different levels of interactions between living organisms in the environment. B) Explain why experiments on different kinds of animals may have relevance for understanding human biology, give also, an example of such approach.

Bradford Assay Fill in the average A595 and A595 of sample minus A595 of blank. Show some sample computation.  Concentration  (micrograms/ml) A595 - trial 1 A595 - trial 2 A595 - trial 3 average A595 A595 of sample minus A595 of blank 0 1.501 1.446 1.447     2 1.624 1.558 1.559     5 1.731 1.749 1.712     10 1.901 1.838 1.892     15 2.161 2.108 2.228     18 2.231 2.277 2.319

Question:- 1b. What does selective toxicity mean in regards to how antibiotics work? INSERT ANSWER

Microbial sensivity lab: Why are Escherichia coli, Pseudomonas aeruginosa, and StaphyLococcus aureus used as standards in the Kirby­ Bauer method?

Briefly explain that what is the important to use immobilization techniques in biosensor?..(explain under5 points)

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281 Customized Subject Test 50. During an experiment, equal aliquots of Escherichia coli and Staphylococcus aureus are separately sonicated at the same level to mechanically disrupt bacterial cells. E. coli cells are disrupted, but there is minimal disruption of S. aureus cells. Which of the following bacterial structures best explains this difference? A) Cytoplasmic membrane B) Outer membrane C) Peptidoglycan layer OD) Phospholipid bilayer OE) Polysaccharide capsule

Explain the basic principles of Elispot Assay

Q2. In one of the experiments, DNA synthesis was inhibited. - what chemical was used to inhibit DNA synthesis, and what is its mechanism of action (how does it inhibit DNA synthesis)? 2. Show the calculations required to make up 250mlof a stock solution of this chemical thất will then be at the working concentration when 1 volume of this solution is added to 10 volumes of cell culture medium.

Exercise 5: Gram-stain: know steps, stains/reagents, and colors of Gm- and Gm+ after each step as well as overall process Predict cell color of Gm - and/or Gm + cells at end of erroneous process Interpret micrographs for Gm reaction