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October 19, 2013 Ligation of CMV + MV9
Part
Concentration
CMV (insert)
15.535 ng/μL
MV9 (vector)
20.838 ng/μL
Calculations
ng insert = bp insert/bp vector * 2 * 50
μL insert = ng insert/concentration insert
μL vector = 50 ng/concentration vector
1
Control
DNA Insert
0.8
0
DNA Vector (MV9)
2.4
2.4
T4 Ligase
1
1
Lign Buffer (2x)
5
5
dH2 O
0.8
1.6
Total 10 µL
10 µL
Bacterial Transformation of CMV Promoter + MV9
Warm 100 μg/mL Amp agar plate at 37°C
Thaw fresh tube of DH5α Turbo cells on ice
Add 30 μL of DH5α Turbo Cells to DNA + dH2 O.
This was also done for transformation of part KAH126
Incubate cells + DNA on ice for 5 minutes.
Label pre-warmed plate.
Transfer cells + DNA onto agar.
Add 8-10 glass beads, shake, then discard beads.
Incubate plates at 37°C overnight and retrieve next day.
