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Proteins
We generally tend to think of proteins only from a dietary lens, as a component of what we eat. However, they are among the most important and abundant organic macromolecules in the human body, with diverse structures and functions. Every cell contains thousands and thousands of proteins, each with specific functions. Some help in the formation of cellular membrane or walls, some help the cell to move, others act as messages or signals and flow seamlessly from one cell to another, carrying information.
Protein Expression
The method by which living organisms synthesize proteins and further modify and regulate them is called protein expression. Protein expression plays a significant role in several types of research and is highly utilized in molecular biology, biochemistry, and protein research laboratories.
Make a conceptual framework based on this concept (Antimicrobial Resistance).
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- Five ml of bacterial culture is added to 45 ml of sterile diluent. From this suspension, the following serial dilutions were made, two 1:100 and one 1:10 dilutions, and 0.1 ml is plated onto Plate Count Agar from the last dilution. After incubation, 186 colonies were counted on the plate. 1. What is the dilution factor, or how much of the original sample was diluted?A sample of water was enumerated using a counting chamber (haemocytometer) and 220 bacterial cells were counted in 25 small squares (each of volume 2.5 x 10-7 cm3). Viable counts were carried out on the same culture using both the pour plate method (incorporating 1 mL samples of a range of dilutions into plate count agar plates) and the Miles & Misra spread plate method, where 0.02 mL samples were spread on sectors of a plate count agar plate. For the pour plate count the average of triplicate samples was 178 colonies per plate of the 10-4dilution. For the Miles and Misra count the average of triplicate samples was 21.3 colonies per sector of the 10-4 dilution. Calculate the total count and the viable pour plate and spread plate counts and suggest possible reasons for the difference between the three different counts.You are given a bacterial culture which has a concentration of approximately 5.0 x 10^8 cells/mL. List a series of dilutions and platings that you could carry out in order to determine the exact concentration of the culture. Note that you must plate four plates from a minimum of two dilution tubes. The volumes plated should be in the range of 0.1 mL – 1.0 mL. Duplicate volumes may not be plated from any one dilution tube. Each plating should aim for a count between 30 and 300 CFUs. You can select any value from 30-300 for CFU and any volume from 0.1-1.0 to find out dilution scheme
- A serial dilution of overnight E.coli culture was performed by pipetting 1ml of a bacterial culture into a 9 ml LB medium. After this, from 10-4 and 10-5 dilution tubes 100µl were plated onto LB agar plates. Upon overnight incubation at 37°C, 200 colonies were counted in 10-4 and 22 colonies were present on 10-5 plates. How many colony-forming units were present per ml of the original culture? If the formula for CFU/ml =no. Of colonies/dilution factor*volume of culture plateFive mL of bacterial culture is added to 45 mL of sterile diluent. From this suspension, the following serial dilutions were made, two 1:100 and one 1:10 dilutions, and 0.1 mL is plated onto Plate Count Agar from the last dilution. After incubation, 186 colonies were counted on the plateTo determine the number of cells in a pure culture of Klebsiella pneumoniae, you have performed a serial dilution using three tubes of sterile saline (9.9 ml each). A sample of 0.1 ml from the culture was added to tube 1. Similarly, 0.1 ml from tube 1 was used to inoculate tube 2, and tube 3 was inoculated using 0.1 ml from tube 2. A nutrient agar plate was then inoculated using 0.1 ml from tube 3 and incubated overnight. The next day, 92 colonies were observed on the plate. How many cfu/ml were in the original culture? Using a Petroff-Hauser counting chamber, the number of cells in the same culture was estimated to be 8.5 • 109 cells/ml. How can you explain these results?
- How do eosin-methylene blue (EMB) agar plates work? What organism(s) are they designed to detect? How would a positive test appear on the plate?Ten grams of hamburger were added to 90 mL of sterile buffer. This was mixed well in a blender. One-tenth of amL of this slurry was added to 9.9 mL of sterile buffer. After thorough mixing, this suspension was further dilutedby successive 1/100 and 1/10 dilutions. One-tenth of a mL of this final solution was plated onto Plate Count agar.After incubation 145 colonies were present. How many colony-forming units were present in the total 10 gramsample of hamburger?What are the possible reasons why culture plates may have too many colonies despite performing serial dilutions and OD600 readings?
- Assuming that serial dilution was carried out in a laboratory experiment in 6 tubes with 9 ml of saline in each tube and 0.1ml of the stock solution was used for initial dilution, pour plate method of inoculation was performed after the dilution process using 1ml of the culture from the samples from the test tubes. It was found out that on the 6th Petri dish after incubation, there was a total of 16 individual colonies counted. Compute the total number of microorganisms present in tube one, assuming that there was no human error in the transferring proces.assuming that serial dilution was carried out in a laboratory experiment in 6 tubes with 9 ml of saline in each tube and 0.1ml of the stock solution was used for initial dilution, pour plate method of inoculation was performed after the dilution process using 1ml of the culture from the samples from the test tubes. It was found out that on the 6th Petri dish after incubation, there was a total of 16 individual colonies counted. Compute the total number of microorganisms present in tube one, assuming that there was no human error in the transferring process.You are given a 1 gram soil sample of unknown bacterial load. After doing 10-fold serial dilutions of the soil in sterile water, 100 uL volumes are taken from each dilution for preparation of pour plates. Following incubation, each half of the 10-8 plate has 46 colonies.a) What was the dilution factor?b) How many bacteria were present in the soil?2. Staphylococcus aureus divides every 20 minutes. A culture begins with 10 bacterial cells.a) After 5 hours, how many generations have occurredb) After 5 hours, how many bacteria are present?3. How many milliliters would you need to prepare a 10-2 dilution from a 10ml starting culture?