Lab Report III (Labs 3-5)Calculate the transformation efficiencies for both ligation reactions:Transformation efficiency = (number of colonies)/(μg DNA)To do this, the amount of DNA that resulted in the number of colonies on each plate needs to be calculated. Remember that not all of the transformation mixture was plated. If no colonies were observed on a plate, calculate what the efficiency would be if there had been one colony.
The efficiency of this transformation must be less than this.Using the molecular weight of the plasmid determined previously, convert the amount of DNA that you used in the transformation of plasmid to the number of moles of DNA. Using Avogadro’s number, convert the number of moles of DNA to the number of molecules of DNA in the transformation.
Assuming that each colony arose from one molecule of DNA, what percentage of the DNA molecules were successfully transformed? What might limit the number of colonies obtained from the transformation?Provide an explanation forany changes in the color of the colonies on the plates for the transformations. Draw all possible ligation products and indicate what color each would be. Explain how the concentration of DNA in the ligation could affect the ratio of the colors observed for the colonies.
Different color variants have been made by altering the amino acid sequences of GFP and RFP. Several of these are listed below. From your results on the absorption and emission spectra, determine which strain of GFP or RFP you have. The peaks will vary slightly depending on the conditions, so find the one that is closest to your observations. Does your gene sequence support this assignment?Variantabsorption maximum emission maximum
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