- Calculate The Concentration Of The Fragments In Lanes 2 5 6 2 W N 1 1kb Dna Ladder 4z E1 4z E4 3 456 Mass Ng 4 1 (60.44 KiB) Viewed 87 times
• Optimized cloning efficiency is 50 ng of vector with 2-foldexcess of insert
• Recommended Insert:Vector Molar Ratio = 2:1
a. Calculate the mass (ng) of insert needed for 50 ng of vectorusing a I:V ratio = 2:1.
Show your calculations, complete the table below, and fill thespaces provided
• Vector length pGEM-4Z = _______ bp
• Insert length Luciferase [LUC] Insert = _______ bp
b. Calculate the volumes (μL) for each fragment [insert& vector] that contain the mass from the question (a)above tobe used in the ligation reaction. Calculate the concentrations ofinsert & vector using the AGE image or nanodrop results of theGel Purification Show your calculations and complete the tablesbelow.
Calculate the concentration of the fragments in lanes 2, 5, & 6 2 W N 1 1Kb DNA Ladder 4Z-E1 4Z-E4 3 456 Mass (ng) --- 43 33 33 40 26 100 Luc-E1 Luc-E2 37 27 33 33 Kilobases 48,- 39= 10 4 Kb Ref. Band 2 kb 8- 6- 5 4 3 2- 1.5- 1- 0.5- 1 2 3 4 5 6 7 Quick Load 1 Kb Extended NEB 50 ng/μL Mass (ng) Kilobases 45 33 33 40 33 26 100 37 27 33 33 BOB O86B43 ||||||| 2- 1.5- 1- 0.5-
Calculate the concentration of the fragments in lanes 2, 5, & 6 1 2 w N 3 456 Mass (ng) Kilobases 48,80 38 10 - 8- 6- 51 1Kb DNA Ladder 4Z-E1 4Z-E4 --- 100 Luc-E1 Luc-E2 化 33 33 4 Kb Ref. Band 2 kb 3 2- 1.5- 1- 0.5- 1 2 3 5 6 7 Mass (ng) Kilobases 45 33 33 40 33 26 100 37 27 33 33 25086543 ||||||||| 2- 1.5- 1- 0.5-