BIO1140-2024-Midterm-Marking scheme-Section B

10) List down 3 different test animals and give samples for which purposes they can be used in testing of a designed biomaterials? 11) What does FDA abbreviation stand for? 12) Order for the below given steps prior to FDA approval ? Controlled clinical trials In vitro testing (“in glass") In vivo testing w/animal models of disease In vivo testing w/healthy animals

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

DATA 24. MA Rollin Hotchkiss and Julius Marmur studied transformation in the ANALYSIS bacterium Streptococcus pneumoniae (R. D. Hotchkiss and J. Marmur. 1954. Proceedings of the National Academy of Sciences of the United States of America 40:55-60). They examined four mutations in this bacterium: penicillin resistance (P), streptomycin resistance (S), sulfanilamide resistance (F), and the ability to use mannitol (M). They extracted DNA from strains of bacteria with different combinations of different mutations and used this DNA to transform wild-type bacterial cells (P* S* F* M*). The results from one of their transformation experiments are shown here: Donor Recipient Transformants Percentage DNA DNA of all cells MSF M* S* F M* SF 4.0 M* S* F 4.0 MS' F 2.6 MSF 0.41 M* SF 0.22 MS* F 0.0058 MSF 0.0071 a. Hotchkiss and Marmur noted that the percentage of cotransformation was higher than would be expected on a random basis. For example, the results show that 2.6% of the cells were…

Experiment: Bradford protein assaygive the answers (4-5 lines) of review questions in the end. the answer should be logical and understandable and without plagiarism. avoid copy-pasting. PLEASE GIVE THE ANSWER OF 3rd AND 4th QUESTION. ITS COMPULSORY.Background information:The Bradford assay is very fast and uses about the same amount of protein as the Lowry assay. It is fairly accurate and samples that are out of range can be retested within minutes. The Bradford is recommended for general use, especially for determining protein content of cell fractions and assessing protein concentrations for gel electrophoresis. The method described below is for a 100 µl sample volume using 5 ml color reagent. It is sensitive to about 5 to 200 micrograms protein, depending on the dye quality. In assays using 5 ml color reagent prepared in lab, the sensitive range is closer to 5 to 100 µg protein. Scale down the volume for the "micro assay procedure," which uses 1 ml cuvettes. Protocols, including…

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

If you wanted the cut time required for centrifugation to half, what would you change? Please write advantages and disadvantages.

Environmental biosensors are analytical devices and the following is a diagram of the structure of environmental biosensors. Briefly describe the working principle of both direct and indirect assays of biosensor taking consideration of any one of the available examples as biological recognition element and physical transducer.

You now redo the experiment but instead of an SDS gel you run a native gel followed by completing the westernblot. Why did you run a native gel? What parameter are you interested in here?

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)

Application Refer to the table below to answer the following items. Base Percentage Source of DNA Adenine Guanine Cytosine Thymine Sea urchin 32.8 17.7 17.3 32.1 Salmon 29.7 20.8 20.4 29.1 Wheat 28.1 21.8 22.7 E. coli 24.7 26.0 Human 30.4 30.1 Ox 29.0 Average %

2. Sanger sequencing question. a). Why the Sanger sequencing chemistry would not work if too much ddNTP is added in the reaction mixtures? if we have too much ddNTP present in our reaction, we would have early termination near their 5' end and we would never generate products that are long enough to determine the sequence in the capillary electrophoresis. b) In a modified Sanger sequencing experiment, products from each reaction were recovered after separation by gel electrophoresis, and their exact molecular masses were determined by mass spectroscopy for the molecular species where all phosphodiesters are ionized. The molecular ions were found as follows: A reaction: 1163, 3050 and 3363; C reaction: 1451 and 3651; G reaction: 1779, 2107, and 2435; T reaction: 2738 and 3954. The reactions used d(AGC) as a primer. What is the sequence of the target DNA? show/explain your work.

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?

Question 3 Listen Which of the following statements describe disadvantages of the classical Sanger sequencing approach when compared to the modern Sanger sequencing method? |Radioactivity must be used, representing an added hazard Fluorescence is used, representing an added hazard Four reaction are used, requiring more reagents and escalating cost OGel electrophoresis is used to separate the various products by size but slowing overall completion of sequencing OA single reaction is used, complicating the analysis O Capillary electrophoresis is used to separate the various products by size but slowing overall completion of sequencing Question 4 4 Listen Electrophoresis of polynucleotides can be done using either agarose or polyacrylamide gels. What is the primary consideration when choosing which polymer gel to use? O Overall charge on polynucleotide sample, greater overall negative charge requires agarose gel with smaller pores to slow the migration of the highly charged sample O…

Please provide explanations with clear work showing how to do the assignment. This is Biochemstry. Please explain each part thoroughly

Procedure A. Applying the terms of science → The following fictional experiment demonstrates the use of the scientific method. Many people in the small Midwestern town of Hootville are stricken with the disease "Buggo." Most of those who get sick recover in seven to nine days. Buggo has been shown to be caused by a bacterium (single for bacteria) called "Gotcha." Antibiotic "X" is a new drug that has been shown to kill Gotcha bacteria in a test tube. Antibiotic X was also found to cure dogs that were sick with Buggo. 3 #m Researchers decided to test the new drug on some of the people in Hootville. They hypothesized that the drug would effectively cure Buggo in humans. They decided that if they gave patients Antibiotic X, the patients would recover more quickly than those who did not take the antibiotic. The researchers prepared tablets containing Antibiotic X. They also prepared a second batch that contained all the same ingredients (inactive ingredients) as Antibiotic X without any…

Question: Summarize The Theory Of SDS-PAGE Electrophoresis.

Gel Electrophoresis Background and Protocol: Gel electrophoresis is a laboratory technique that separates molecules by size using an electric current.  The test has a positive and negative side.  Do you believe DNA should be loaded on the positive (red) side or the negative (black) side?  Please explain why using scientific reasoning. Will larger DNA bands be closer or farther away from the well where you administered samples?  Why?

Please answer   Resolution (R) is a term used to say something about how good separation you have in a chromatographic system. What experimental conditions can you change to make a bad separation better? Explain.

Using a schematic diagram, summarize the following steps in preparing competent cells for transformation: Inoculate a single colony of E. coli into 5 ml LB broth and incubate overnight at 37°C with moderate shaking (250 rpm). Add 200 μl of the culture into 50 ml LB broth and incubate overnight at 37°C with moderate shaking (250 rpm) to an OD600 = 1.3 to 1.5. Aliquot culture into five 15-ml pre-chilled, conical tubes. Leave tube on ice 5 to 10 min. Centrifuge cells 7 min at 1,600 × g (3,000 rpm), 4°C. Pour off supernatant and resuspend each pellet in 10 ml ice-cold CaCl2 solution (50 mM CaCl2), perform resuspension very gently, and keep on ice. Centrifuge cells 5 min at 1,100 × g (2,500 rpm), 4°C. Discard supernatant and resuspend each pellet in 10 ml ice-cold CaCl2 solution. Keep resuspended on ice for 30 min. Centrifuge cells 5 min at 1,100 × g, 4°C. Discard supernatant and resuspend each pellet in 10 ml ice-cold CaCl2 solution. Dispense cells (250 μl) into pre-chilled, sterile…

f) Examine the advantage of DNA based biosensor as compared to enzyme biosensor?

question: Can you summarize and explain for me what you want to tell in the article below? When I read it myself, I do not understand exactly what is meant by the article. It would be nice if you could highlight the important points. You can use them in a figure or diagram to explain. thank you and hava a nice day :) Article: Biomimetic Engineering of Nanodelivery Systems: Artificial Viruses in the Making In an effort to engineer the next generation of nanoscale vectors, scientists have moved from using inorganic components aimed at obtaining inert structures to utilizing biological building blocks that are able to convey additional functionalities to the resulting construct. To cope with the complexity of the body and to evade the multiple layers of defense that tissues and organs have, it is critical to rely on the ability of certain materials to interact with, rather than to eschew, the biology of our body. Every NP system conceived to date faces one common fate: whether injected,…

Give information about the microchip that could control drugs invented by Robert Langer and Michael Cima. When the studies started, how did the idea come out, who found it, which university the studies were carried out, what exactly does the product work, when was the patent obtained? Write important information about this polymer wafer (Please don't give short answer)

Results: Both nonencapsulated and encapsulated bacteria grow. Procedure II: Extract made from dead encapsulated cells treated with protein-degrading enzymes before adding extract to culture medium. Nonencapsulated bacteria added to culture medium. Results: Both nonencapsulated and encapsulated bacteria grow. Procedure III: Extract made from dead encapsulated cells treated with DNAse (an enzyme that selectively destroys DNA) before adding extract to culture medium. Nonencapsulated bacteria added to culture medium. Results: Only nonencapsulated bacteria grow. 14. A reasonable conclusion to draw from the results of the experiment is that (A) DNA is the genetic material (B) DNA replication is semiconservative (C) DNA is a double helix (D) DNA is translated into protein (E) mutation is a change in the genetic material 15. What was the purpose of treating the extract with protein-degrading enzymes in Procedure II? (A) To demonstrate that the transforming factor is an enzyme (B) To…

With detail, compare and contrast the following 5 real-time assays; Taqman, SYBR Green, Molecular Beacon, FRET, and Scorpion. Please write complete sentences in paragraph form. Identify how the assays are similar but more importantly, what are the distinguishing features of each of the technologies.

BIOTECHNOLGY Date: Name: Instructor: Section/Group:. POST-LAB QUESTIONS 1. In one or two sentences, summarize the technique of gel electrophoresis. 2. How does the process of gel electrophoresis separate DNA fragments? 3. Why is the fact that DNA has a negative charge so important in the gel electrophoresis process? Biotechnology 165

DNA Extraction by Alkaline Lysis Procedure: 1. Spin 1.5 ml of cells in a microcentrifuge at maximum speed (12,000 rpm) for 20s to pellet. Remove the supernatant completely with a Pasteur pipet or a plastic pipettor tip. The spins can be performed at 4C or at room temperature. Longer spins make it difficult to resuspend cells. 2. Resuspend pellet in 100µl GTE solution and let sit 5 min at room temperature. Be sure cells are completely resuspended. 3. Add 200µl NaOH/SDS solution, mix by tapping tube with finger, and place on ice for 5 min. 4. Add 150µl potassium acetate solution and vortex at maximum speed for 2s to mix. Place on ice for 5-15 min. Be sure mixing is complete. 5. Spin 3 min at 12,000 rpm to pellet cell debris and chromosomal DNA. 6. Transfer 0.4 ml supernatant to a fresh tube, mix it with 0.8 ml of 95% ethanol or 0.4 ml isopropanol, and let sit 2 min at room temperature precipitate nucleic acids. 7. Spin at 12,000rpm for 3 min at room temperature to pellet plasmid DNA and…

briefly describe the working principle of both direct and indirect assays of biosensor taking consideration of any one of the available examples as biological recognition element and physical transducer

DNA Isolation A. What is “cell lysis” and why would you want to lyse cells when doing a DNA extraction? B. What do the “proteases” like meat tenderizer do to the DNA? C. What two ingredients help precipitate the DNA out of solution?

Drug substances are useful based on their biological activity. Thus, an assay must be developed to ensure that this important function is preserved during purification. There are three major types of biological activity assays including animal model assays, cell-line-derived assays, and in vitro biochemical assays. Please find an example of each (in literature, online, etc.) and briefly Include a reference for each (no particular reference format required, can simply include a web link if easiest).

a. For the E.coli haploid genotype below, determine the phenotype for the following proteins. Assume that any genes or structural elements not shown are wildtype. lacls lacz lacY* lacz [ Select ] lacY [ Select ]

8:25 Screenshot_2021120 What I Can Do Activity 6. What is your stand? Directions. Below are some of the arguments about the use of transgenie organism. In your own perspective, explain your answer in not more than 5 sentences. 1. Among the cited examples of GMO, which do you think is the most beneficial? 2. If you are a farmer would you take the chance of growing crops that are pest resistant? Why or why not? 3. Considering the knowledge gained genetic engineering, would you try to patronize GMO fruits and vegetables? Why or why not? 4. Is creating or altering genes of an organism a form of Blasphemy to the creator (God)? Why? 5. Is genetic engineering morally permissible or not?

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

Remember that although there are many interesting ideas about genetic engineering of plants and animals, this is specifically about GE bacteria. Please be sure you are answering the following questions    3) What are the benefits (or potential benefits) of the engineered bacterium?     4) What are the risks (or potential risks) of the engineered bacterium?

Please offer a clear explanation of the ChIP assay.  A. What characteristics of these cells are being assessed? Why would this be valuable to a research team? B. Define assay and explain how samples are managed step by step with a clear explanation of each step. C. What type of data is being generated by this experiment?

The Nicotinamide Adenine Dinucleotide (NAD) Assay (SIGMA Kit MAKO37) requires preparation of a Standard curve: the first time the blanks are run. each time the assay is run. the first time the assay is run. each time the blank is run.

Droplet based single cell DNA sequencing uses a microfluidic platform which generates droplets encapsulating individual cells. A micrograph of such a device [Zilionis et al, Nature Protocols 2016] is presented in Figure Q3. The channels are 25 μm deep. (a) (b) Oil Figure Q3. Micrograph of the microfluidic droplet device. Scale bars are 100 µm. Adapted from Zilionis et al, Nature Protocols 2016. Red arrowheads show individual cells, and black arrows indicate flow direction. (c) RT/lysis reagents Cells Barcoding hydrogel beads A company wants to explore the design of a prototype of the device made from PDMS. Describe a suitable fabrication process, explaining the rationale behind each individual step. Schematics can be drawn to illustrate your response. You may want to refer to the datasheets at the end of the script. Testing of the prototype was successful and the company wants to explore the feasibility of manufacturing the device from glass. Describe the fabrication process for this…