1 GENERAL BIOLOGY LAB 1 (BSC1010L) Lab #10: Biotechnology ...

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4. Record your results Table 2.Table 2:ConditionLB -Results(growth/no growth)Type of GrowthObserved(lawn/colonial)Did your results agree with yourpredictions in Table 1?(if not, provide a possibleexplanation)LB/Amp -LB +LB/Amp +5. Calculate transformation efficiency.Transformation efficiency is a measure of how successful host cells are in uptaking theplasmid, where uptake is expressed as the number of transformed colonies per µg ofplasmid DNA.Note: For this exercise, you will need to count the number of colonies present on yourLB/Amp+ plate. Follow steps a through e to calculate the transformation efficiency ofyour E.coli strain.a. The total amount (µg) of plasmid DNA used can be calculated with the followingformula.µg DNA = concentration (µg/µL) of DNA x volume of DNA (µL)b. Calculate the total volume of cell suspension prepared in the + DNA tube.Total volume (µL) = amount (µL) of CaCl 2 + amount (µL) of plasmid + amount (µL) of LB10
c. Since only a portion of the + DNA solution was added to the LB/Amp + plate, youwill need to calculate the fraction of DNA spread onto this plate using the formulabelow:Fraction of DNA spread = Volume (µL) spread on LB/Amp+ plateTotal sample volume (µL) in + DNA tubed. Using the values obtained for parts a and c, determine the actual amount of DNA (µg)present on your plate.Total amount (µg) of DNA = µg of DNA x Fraction of DNA spreade. Calculate transformation efficiency.Transformation Efficiency = Total number of cells on the LB/Amp+ plate__Total amount of DNA spread on LB/Amp+ plateQuestions:1. What role did the following compounds or steps play in bacterial transformation?a. Calcium Chlorideb. Heat shockc. Agard. LB Brothe. Ampicillin11
Page 3 and 4: Table 1:LB -Condition Expected resu
Page 5 and 6: Figure 17. Return the + DNA tube to
Page 7 and 8: 17. Using a sterile transfer pipett
Page 9: A. B.Figure 6. Plates illustrating
Page 13: 5. Can you think of a case where th

1 GENERAL BIOLOGY LAB 1 (BSC1010L) Lab #10: Biotechnology ...

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