Calculation of the Transformation Efficiency The efficiency of transformation in

Question

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 270 colonies on the L-agar + kanamycin plate with 0.10 ml of the undiluted suspension. You see 180 colonies on the L-agar plate with 0.10 ml of the 10-7 dilution of the suspension. What is the transformation frequency?

Explanation / Answer

Transformation efficiency = (conc. of transformants)/(conc. of recipient cells)-----------------------1

conc. of transformants = (number of colonies on selective agar plate) x (Dilution Factor) / 0.10 ml-----------------------2

(conc. of recipient cells) = (number of colonies on L-agar plate) x (Dilution Factor) / 0.10 ml--------------------3

All the formulas given in question, we have to calculate transformation efficiency in bacterial cell.

First we have to calculate concentration of transformats- by counting visible number of colony on selective agar plate- 270 (L-agar + kanamycin plate), now put this value in formula number 2,

conc. of transformants = (270) x (00) / 0.10 ml = 2700-------------------------4

now we have to calculate concentration of recipient cells by counting visible number of colony on L-agar plate- 180 and here given range of dilution factor 10-7, we take 10 as a dilution factor. Now calculate

conc. of recipient cells = (180) x (10) / 0.10 ml = 18000--------------------------5

now we have concentration of both transfrmat or recipient cells. Put this value in equation first

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

Transformation efficiency = (2700)/(18000)

Transformation efficiency = 0.15

So the transformation frequency of bacterial cells = (no. of transformed cell)/total number of transformed cells.

Transformed colony = 270 and total number of transformed cell = 450

So

transformation frequency= (270)/(450) = 0.6 or 60%

the frequency of transformation is 60%

Related Questions

Navigate

• Browse (All)
• Browse C
• Subjects

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.