Essay About Basic Cell Culture Techniques And 8Basic Cell Culture Techniques Name

Basic Cell Culture Techniques.
BIOL 4380 Lab Report 8Basic Cell Culture Techniques Name: Katherine Reittinger  Lab Section: 304  Instructor: Dr. Irina Borovkov Date: March 28, 2019 Experiment #8 Lab ReportBasic Cell Culture TechniquesThis report is worth 30 points, must be typed, and is due at your next scheduled lab period.  Please re-type the question before typing your answer.  Calculations, formulas, tables, and graphs may be hand written.  The term “publishable” means that all the necessary components (including a figure caption) required for publication in a scientific journal are present and correct.  1. (8 points) a. What was the overall goal of our series of experiments (3, 4, 5, 6, 8)? (3 points)        The overall goal of this series of experiments was to determine if our bacterial gene         of interest can be expressed in mammalian cells if under a CMV promoter.b. Why did we transfect pTUDβ we constructed into mammalian cells? What was the purpose of transfecting the cells with pTETbeta? (3 points)pTUDbeta is our recombinant plasmid that contains the beta-galactidosase ORF and the CV promoter. We transfect it into mammalian cells to be able to check for beta-galactidosase activity. Transfecting mammalian cells with pTETbeta acts as a control. This transfection should result in no blue cells post staining for beta-glalactidosase activity.c. Did we get an answer to our questions? Explain. (1 point)Yes. After staining our transfected HEK293T cells with X-gal staining solution we were able to observe some blue cells. This indicates the beta-galactidosase in pTUDbeta was expressed.d. What is your estimate of the percentage of HEK293T cells that became transfected with the pTUDβ plasmid? (1 point)        About 40% of the mammalian cells were transfected with pTUDbeta.2. (4 points) Report the titer (cells/ml) of  cells in the two dishes that your group counted.        Plate 1Plate 2Original count107.25 cells (avg.)144.25 cells (avg.)Cells per ml1,072,500 cells/mL1,442,500 cells/mLTotal number of cells in suspension8,580,000 cells (total volume = 8mL)11,540,000 cells(total volume = 8mL)Were there differences in cell count between your and your partner’s plates? Explain why there may be differences in the data for plate 1 and plate 2.        Yes, there are differences. Mainly, the cell confluency was higher for plate 2 than         plate 1.1 pt What is the approximate diameter of mammalian cells in suspension, estimated from observing the cells in the hemocytometer?  (Remember, one of the nine large grids on the hemocytometer has the dimensions of 1mm x 1 mm.)