Question

8. You have two plasmid preparations "A and "B" (illustrated below) each at concentrations of 100nguL. You wish to subclone a 1.5 Kbp EcoRI fragment from plasmid·A" into the unique EcoRI site of a 3.0Kbp vector"B". EcoRI 1.5 Kbp Total size 3.0Kbp BT Total size 7.5Kbp Complete the following table to indicate the volumes of each ingredient required for the preparation of a 50μL ligation mixture containing 50ng of vector and an amount of digested plasmid A required to have an insert to vector ratio of 3:1. Ingredient Volume (μL) EcoRI digested plasmid 50ng EcoRI digested Plasmid "B* 5X ligation buffer T4 DNA ligase Water 10

See image

please show full solution, (Indicate your answers to two digits after the decimal)

Answer 1

Ingredient	Volume (uL)
EcoRI digested plamsid A	1.5
50ng EcoRI digested plasmid B	0.5
5X ligation buffer	10
T4 DNA ligase	1
Water	37

Plasmid A stock concentartion is 100ng/ul we need 150ng so 1.5ul

Plasmid B stock concentration is 100ng/ul we need 50ng so 0.5ul