Consider the following image of a stained agarose gel representing the separation of a plece of DNA that has been digested by a series of restriction enzymes. Assume no digest fragment is of identical size to another digest fragment. nucleotide pairs (kb) 8 S 45 3.5 AA size Hindill + markers EcoRI Hindill EcoRI - 1st attempt Based on all data presented in the stained agarose gel, which of the following statements is correct? Choose one: OA. The original piece of DNA was circular and was cut by EcoRI twice and Hindill only once. OB. The original piece of DNA was linear and was cut by EcoRI twice and Hindill only once. OC. The original piece of DNA was circular and was cut by EcoRI once and not at all by Hindi!!.

DNA samples from four individuals were cleaved with the same MW restriction endonuclease. The DNA fragments were separated by gel clectrophoresis, transferred to a membrane, and hybridized with a 12 kb 10 kb DNA probe complementary to a region between sites C and D (see 8 kb hybridization line). The image of the southern blot shows the labeled DNA bands and 6 kb molecular weight (MW) markers. The lane labels I, II, III, and IV -5 kb correspond to individuals I, II, III, and IV. Assume that fragments such as C-D and C-E are clearly resolved in this gel system. Fragment sizes are as given: A-B is 4 kb, B-C is I kb, C-D is 5 kb, and D-E is 650 bp. Individual I has five cleavage sites (A, B, C, D, and E) for the restriction endonuclease. DNA homologous to probe Which individual has at least one point mutation that eliminates restriction site C only? O II IV cannot be determined II Which individual has at least one point mutation that climinates restriction sites B and C? III O IV cannot be…

How many fragments and how big each fragment should be after digesting the plasmid with BamHI? How many fragments and how big each fragment should be after digesting the plasmid with PvuI? How many fragments and how big each fragment should be after digesting with both enzymes? Show your calculation/explanation.

How much 340bp insert DNA should be added to a ligation in which 50ng of 4.3kb vector will be used? The desired vector: insert molar ratio needs to be 1:3.