Background Calculating Transformation Efficiency Restriction digest & ligation: You add restriction enzymesto 0.05ug of each plasmid in separate pAMP and pKAN tubes tubes andheat the restriction enzymes by placing them in a hot water bathfor 20 minutes. Afterwards, you label 1 tube pAMP/pKAN and combine4 ul of digested pAMP, 4 ul of digested pKAN, 1 ul of ligase, and 1ul of ligase buffer and leave it to incubate overnight. You thenlabel an empty tube -pKAN/-pAMP which will be used as your control(no plasmids). Bacterial Transformation: To the 2 microcentrifuge tubes labeled+pKAN/+pAMP and -pKAN/-pAMP you add 250 µl of transformationsolution (0.05M CaCl₂). Using aseptic techniques you transferbacteria colonies into the -pKAN/-pAMP and +pKAN/+pAMP tubes makingsure to avoid cross contamination. You incubate the tubes for 10minutes in ice, transfer them to a water bath for 50 seconds, andthen back in ice for 2 minutes. You then pipette 250 uL broth intoeach tube and incubate them for 10 minutes to make the bacteriahappy again. Afterwards, you pipette 100 ul of each transformationmixture and spread it onto the appropriately labeled agar plates.You parafilm and incubate the plates of E. coli overnight. The nextday you got the following results.   Plate Label # of colonies - LB agar Too many to count (TMTC) - LB pAMP/pKAN No growth + LB agar TMTC +LB pAMP/pKAN 2 The questions that I need help on: Calculate the transformation efficiency in CFU/ug of DNA fromthe +LB/AMP/KAN plate. The concentration of the pKAN/pAMP is 0.1ug/ul. The amount of plasmid DNA spread on the plate was 1 ug. Determine the number of colonies growing on theLB/amp/kan agar plate based on the informationprovided. Determine the amount of plasmid DNA (in ug) spread onthe LB/amp/kan agar plate from the informationprovided. DNA spread (ug)= Volume spread on plate (ul) X DNA intransformation (0.05 ug)                                            Totalvolume of transformation                                        (addup all the highlighted volumes) Calculate the transformation efficiency (CFU/ug) bydividing up the number of colonies on the plate by the amount ofDNA spread on the plate. Basically divide the answer from #1 byyour answer to #2