Thursday, December 26, 2013

Lesson 4: Extension Activity, Calculate Transformation Efficiency

 (Athira)
  1. Determining the Total Number of Green Fluorescent Cells

    Total number of colonies = 41
  2. 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 =
    41
    Micrograms of pGLO DNA spread on the plates
    0.16




    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

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

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