- Determining the Total Number of Green Fluorescent Cells
Total number of colonies = 41 - Determining the Amount of pGLO DNA in the Bacterial Cells Spread on the LB/amp/ara Plate
a. Determining the Total Amount of pGLO plasmid DNA
Total # of pGLO DNA (μg) = 0.08 x 10 = 0.8 (used to calculate transformation efficiency)
b. Determining the fraction of pGLO plasmid DNA (in the bacteria) that actually got spread onto the LB/amp/ara plate
Fraction of DNA = 100/510 ~ 10/51 (used to calculate transformation efficiency)
pGLO DNA spread (μg)= 0.8 x 10/51 = 0.15682745 μg
# of colonies on LB/amp/ara plate =41Micrograms of pGLO DNA spread on the plates0.16
Transformation efficiency= 41/0.16= 256.25 transformants/ μg
Analysis
How would scientists
report 10,000 transformants/ μg in scientific notation?
1.0 x
104 transformants/ μg
How would scientists
report 40,000 transformants/ μg
in scientific notation?
4.0 x 104 transformants/ μg
How would scientists
report 960,000 transformants/ μg
in scientific notation?
9.6 x 105 transformants/
μg
Report your calculated
transformation efficiency in scientific notation.
1.56 x 102 transformants/ μg
1.56 x 102 transformants/ μg
Use a sentence or two
to explain what your calculation of transformation efficiency means.It means
the # of E.coli bacteria that efficiently
transformed or had the gene of interest.
How does your
transformation compare to the general agreement set by biotechnologists?
#
of colonies on LB/amp/ara plate =
|
41
|
Micrograms
of pGLO DNA spread on the plates
|
0.16
|
We did not reach their agreement, we are under by about 543 transformants/μg.
Fill in the following chart and show your calculations to your teacher:
# of colonies on
LB/amp/ara plate =
227
|
Micrograms of DNA
spread on the plates =
0.157 μg
|
Transformation
efficiency =
1445.86 transformants/μg
|
0.08 x 10= 0.8 μg
100/510 x 0.8 = 0.157 μg
227/0.157 = 1445.86 transformants/μg
Extra Credit Challenge
300/0.157 = 471 colonies of bacteria
