Picture this: You are on the verge of completing a key experiment for your PhD dissertatic project... and you realize you are running short on cell culture medium. To prepare a new 500 mL bottle, you are given the following recipe: Please complete the table by filling in the needed volumes to achieve the IMDM with 10% heat inactivated fetal bovine serum (HI FBS): IMDM 500 mL HI FBS (100 %) A mL 2 mM L-glutamine (200 mM) B mL 1,000 units-mg Pen/Strep C mL (10,000 units Pen/Strep per mL) *Please note: stock concentrations are indicated in blue and desired in black. A = mL B = mL ml
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- Consider a batch culture of sphere-shaped bacteria in a growth medium or broth, in which the mean cell diameter of the bacteria is 2.0 μm (micron). Show all your calculations and assumptions in answering the questions below: What is the volume of one of these sphere-shaped cells, expressed in liters? If there are 3 x 10^12 cells in one liter of this culture broth, what percentage of the volume is occupied by bacteria and what percentage is occupied by the cell culture broth (water only)? If the cells contain 80% water, what is the dry cell concentration, expressed as “grams dry weight/Liter of broth”? You should assume that the density of the cells dry mass is approximately 1.0 gr/cm3For most cell types, serum must be added to the media to get cells to grow and proliferate in culture. Nevertheless, serum is very expensive and can complicate experiments utilizing the cells. Many companies have developed "artificial serum" which enables cells to multiply without the use of serum. See data below. What are likely components of the artificial serum that enables cells to grow. Explain (limit 3-4sentences). Non 2% Human plasma 1% Artificial serum(XF) 1% Artificial serum(AF) 104 cells/cm² Cell density cultured after 4 days 127 10- 2 0 NON 2% 1% 1% Human Artificial Artificial plasma serum(XF) serum(AF) PDL [times] 30- 25+ 20- 15- 10- 5- 0 -2% Human plasma 0 Subculture 1% Artificial serum (XF) 1% Artificial serum(AF) 5 10 15 20 25 30 Culture days Medium: HFDM-1 (+) was used.A Manton-Gaulin homogenizer is used for cell lysis. Which of the following are true? Increasing the number of passes can increase fine cell debris and make subsequent clarification more difficult. Cell growth conditions and phase of growth at which cells are processed can impact cell disruption. Decreasing temperature can increase protein release. Decreasing pressure reduces the number of passes required. Explain what is meant by the extent of cell disruption is 0.9.
- Two co-op students at your start-up company have been asked to evaluate the rate of reaction occurring in a transparent gel particle containing immobilized mouse melanoma cells. The equation for the reaction rate ris is: Vmax T'As Km+CAs where R=3.2 mm, Vmax= 0.12 gmol s' m3, CAS = 41 gmol m3, and Km=0.8 gmol m3. One student reports a reaction rate of 1.6x10 gmol st; the other reports 1.6x10 1º gmol s1. You left your calculator on the bus this morning, but must know quickly which student is correct. Use an order-of- magnitude calculation to identify the right answer. Which student is correct?Propidium iodide (PI) can be used to study both cell division and apoptosis using flow cytometry. Describe how PI is used to study the two cellular events with the principles explained. You are expected to give the full principle of the flow cytometric analysis of apoptosis.During incubation, prepare dilutions of the standard antiserum which constitutes the standard curve for the assay (concentration in ng/mL). Add 500 µL of PBS-milk to the supplied microtube containing 500 µL of standard antiserum to obtain Standard 1 [500 ng/mL]. Identify seven microtubes for standards 2 to 7 and place 500 µL of PBS-milk in each. Calculate how much of antiserum is used in each standard ?
- Glucose 6-Pase is an enzyme present in liver cells but not in muscle cells. The enzyme is bound to the endoplasmic reticulum which forms microsomes when the cells are hydrolyzed. Which of the following is the first critical step in isolating the glucose 6-Pase? A. Preparing the appropriate buffer for isolating the enzyme. B. Identifying the tissue from which to isolate the enzyme of interest and from which additional purification techniques can be applied. O C. Determining the appropriate method or reagent to lyse the cell O D. Obtaining the fraction from the cell which probably contains the enzyme. E. Determining the appropriate chromatographic method to remove contaminating proteins or other biomoleculesImmunofluorescence and Fluorescent livecell imaging techniques can both be used to determine protein localisation.List the advantages and disadvantages of using immunofluorescence andfluorescent live cell imagingMs. Michi is a researcher in a renowned pharmaceutical company based in Singapore. One of the cell lines in the lab is suspected to be contaminated. She is culturing NIH 3T3 mouse embryo fibroblast cell line (NIH/3T3 (ATCC® CRL-1658TM). Figure 1 below illustrates her observation under the inverted phase contrast microscope. a) b) c) d) e) Figure 1 (Routray et al., 2016) What type of contamination is her laboratory facing? Explain your answer. Give FOUR (4) suggestions to Michi in order to prevent such contamination in future Besides the contamination stated in Question 2 a), name FIVE (5) other contaminations that Michi should be aware of. During a routine check, Michi's superior requests her to characterize cell lines available in the storage to determine their authenticity. Describe TWO (2) effective cell line characterization methods that she can utilize for this purpose. The pharmaceutical company wants to improve its toxicity testing by replacing monolayer culture with spheroid…
- The ability of cells suspended in soft (semi-solid) agar medium to form colonies is a good predictor of which of the following? (select all that apply)? A. The cells have lost contact inhibition B. These cells will form tumors if injected subcutaneously into immunocompromised mice. C. These cells have stopped proliferating and will die. D. These cells have an increased requirement for growth factors. E. These cells would fail to grow in normal medium.For the study of alanine production by a recombinant strain of E. coli, cultivation was carried out in a benchtop bioreactor with 4.5 L of culture medium, using glucose as a limiting substrate. During the cultivation, there was no lag phase and the cells showed exponential growth for 5 hours. The following table presents the results of the analysis of ammonia and glucose consumption, and alanine accumulation throughout the cultivation. Knowing that 500 mL of a cell suspension at a concentration of 5.0 g/L (inoculum) was added to the 4.5 L of medium in the reactor and that the YX/NH3 previously determined was 7.5, calculate:a) the maximum specific growth rateb) YX/S and YP/S yield factorsc) How long would it take to reach Cx = 30 g/L if the cells continued with the exponential growth profile until the end of the culture (without nutrient deprivation or any type of inhibition)?d) Describe how the mathematical treatment of the data should be done to determine the type of product formation…https://studylib.net/doc/8245959/lab-7--got-milk%3F follow and open the link then answer the question number 4: Bacteria grown solely on lactose will grow, as will bacteria grow only on ONPG. However, cells grown solely on TMG will not grow, and will instead eventually die. Based on the structures provided below, why are cells capable of growing on ONPG and lactose, but not TMG alone?