ANFS310 Cloning & Transformation Data Analysis.docx - Cloning and Transformation Data Analysis Stevens Due May 8 2019 for Wednesday sections

ANFS310 Cloning & Transformation Data Analysis.docx -...

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Cloning and Transformation Data Analysis Name: Jordyn Stevens Due May 8, 2019 for Wednesday sections Due: May 9, 2019 for the Thursday sections 1. Fill in the table below with your data. Determine transformation efficiency (TE) of the plasmid (pUC8) into E.coli JM109, based on the total number of transformed lacZ- colonies on the ligation plate. Show your calculation for the transformation efficiency (TE). The formula for transformation efficiency is below. All the numbers needed for the formula are in your handout. TE = number of transformed colonies mass of plasmid mass of plasmid = (concentration of plasmid) x (volume of plasmid put in your tube) x (volume of suspension of plasmid spread/total volume of suspension) For total volume of suspension do not count the 15 uL of plasmid (transferred from the original ligation tube to the new ligation tube in transformation) because it is counted in the first part of the equation. Mass of plasmid : Ligation: Mass of plasmid = (0.006 µg/µl) (20 µl) x (100 µl/500 µl) 0.12 x 0.2 = 0.024 = 2.4 x 10 -2 Control: Mass of plasmid = (0.004 µg/µl)(20 µl) x (100 µl/500 µl) 0.08 x 0.2 = 0.016 = 1.6 x 10 -2 TE : Ligation: TE = (84/0.024) = 3.5 x 10 3 cells/µg Control: TE = (1/0.016) = 6.25 x 10 1 cells/µg
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2. Draw a cartoon that depicts an E. coli cell transformed with pUC8 resulting in a functional lac Z gene. Indicate the effect of this cell on the surrounding
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  • Summer '19
  • DNA, Transformation Data Analysis

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