Cells are kept in suspension in RPMI a media for glowing cells. You have 30 mL o
ID: 481154 • Letter: C
Question
Cells are kept in suspension in RPMI a media for glowing cells. You have 30 mL of a cell suspension at a concentration of 7.2 times 10^6 cells/mL. You need to dilute a portion of these cells such that 100 mu L of the new suspension will deliver 1 times 10^5 cells when plated in the wells of a 96-well microstate plate. How would you dilute some cells so you could prepare 48 such wells? Your answer should account for 5-25% extra volume as a pipe ting allowance (answers may vary). ______ mL cells _______ mL RPMI _______ Total volume of cells prepared It would be much appreciated if you can help me with the recipe of the buffer above. Thank you.Explanation / Answer
Total cells in the stock suspension=30ml*(7.2*10^6 cells/ml)=216*10^6 cells
To prepare 48 wells number of cells required=1*10^5 cells/well* 48=48*10^5 cells
volume needed to get 48*10^5 cells =48*10^5 cells/7.2*10^6 cells/ml=6.7*10^-1ml=0.7 ml
So 0.7ml of stock is to be taken for dilution
Now V1=volume of stock cell suspension=0.7ml
C1=7.2*10^6 cells/ml
C2=concentration of wells=10^6 cells/ml [1*10^5 cells/100uL=1*10^5 cells/100*10^-6 L*10^3ml/L=1*10^5 cells/0.1ml=10^6cells/ml]
V2=volume of diluted suspension taken for each well
V2=C1 *V1/C2*48=(7.2*10^6 cells/ml)*0.7ml/48*(10^6 cells/ml)=0.1 ml(volume for each well)
Hence 0.7 ml stock is to be diluted to 0.1*48=4.8ml
total volume of cells prepared=4.8ml
0.7 ml stock cells
4.1 ml RPMI added to stock
Related Questions
drjack9650@gmail.com
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.