8. Plasmid was isolated from bacterial cells and was subjected to restriction digestion using three different restriction enzymes as shown in the gel below. Standards are in Lane 5 and the uncut control is in Lane 1, and roughly the same amount of total DNA in sample lanes 1-4. Considering the given data answer the questions below.
i) Give the order of the DNA conformation in lane 1 from top to bottom?
a. Opencircular, supercoiled and linear.
b. Supercoiled, opencircular and linear.
c. Linear, supercoiled and opencircular.
d. Linear, opencircular and supercoiled.
ii). Approximately how big is your original plasmid?
a. 10 kb
b. 3.5 kb
c. 6.5 kb
d. 9.5 kb
iii) How many times did the enzyme cut in Lane 2 digest the plasmid?
a. 1
b. 2
c. 3
d. 4
iv) How many times did the enzyme used in Lane 3 digest the plasmid? Does the data seem reasonable?
a. 2 times and the data are reasonable.
b. 1time and the data are reasonable.
c. 2 times and the data are not reasonable.
d. 1 time and the data are not reasonable.
v). What are some likely explanations for the smearing detected in Lane 4?
a. Due to absence of DNA
b. Due to DNA degradation
c. Due to restriction digestion
d. Reason B or A
Answers to question 1-C; 2-D; 3-B; 4-D; 5-C; 6-B; 7-D
Answers to question 8: i-D; II-C; iii-B; iv-C; v-D
