1. You prepared a dilution 1 in 2 of trypsinized CHO-K1 cells. Then, you took an aliquot of the cell suspension and counted the cells using a hemocytometer. There were 75 cells in the 16 little squares. Please explain how to get each!

a. Calculate the concentration of the cells prior to the dilution. Show your calculations. Use scientific notation to express the result.

b. Now, you want to seed a 25cm² flask with 750,000 cells. Calculate how many ml of the original cell suspension you will use

c. If the final volume of incubation is 4ml. How many ml of complete medium you will need to add?

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Tod ThielLv2

28 Jun 2019

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1. You prepared a dilution 1 in 2 of trypsinized CHO-K1 cells. Then, you took an aliquot of the cell suspension and counted the cells using a hemocytometer. There were 75 cells in the 16 little squares.

a. Calculate the concentration of the cells prior to the dilution. Show your calculations. Use scientific notation to express the result.

b. Now, you want to seed a 25cm2 flask with 750,000 cells. Calculate how many ml of the original cell suspension you will use

c. If the final volume of incubation is 4ml. How many ml of complete medium you will need to add?

Two T-75 flasks containing anchorage-dependent cultures were trypsinized. The cells from both flasks were pooled, and the resulting cell suspension was transferred to a conical tube. The total volume of liquid cells suspended in media was 20.5 mls. Although these cells are routinely split 1:2, anyone handling these cultures is requested to assess the viability of the population. 0.5 ml of the cell suspension was added to a tube containing an equal volume of trypan blue, the tube was agitated and hemocytometer was loaded with the resulting suspension. Both grids were covered with appropriate amount of cell suspension, and the following counts were obtained. Grid #1: 5 squares counted for a total of 176 cells (5 out of 176 cells were stained blue) Grid #2: 5 squares counted for a total of 195 cells. (5 out of 195 cells were stained blue) What is the average number of viable cell per square? What is the % viability? What is the total number of cells available for subculturing? Since these cells are routinely split 1:2, approximately how many millions of cells will be used to seed each of the 4 flasks?

ceruleanclam711

URGENT - Due in 1 hour! Please help!

This is a numerical response problem.

Calculation of the Transformation Efficiency
The efficiency of transformation in a bacterial conjugation experiment is defined as:

(Transformation efficiency) = (conc. of transformants)/(conc. of recipient cells)

To calculate the concentration of transformants you use the viable plate count formula and the data from the selective agar plates:

(conc. of transformants) = (number of colonies on selective agar plate) x (Dilution Factor) / 0.10 ml

To calculate the concentration of recipient cells you use the viable plate count formula and the data from the non-selective agar plates:

(conc. of recipient cells) = (number of colonies on L-agar plate) x (Dilution Factor) / 0.10 ml

Data
You see 240 colonies on the L-agar + kanamycin plate with 0.10 ml of the undiluted suspension.
You see 240 colonies on the L-agar plate with 0.10 ml of the 10^-7 dilution of the suspension.
What is the transformation frequency?

(Express your answer using 3 significant figures and scientific notation in a format the computer recognizes.
For Example: If your calculations give you an answer of 8.1818181818 x 10^-8, enter it as: 8.18e-8.)
(If your answer is within 5% of the correct value you will recieve credit for the problem.)