# Haynes Lab:Notebook/Engineering PC-TFs/2013/10/19

(Difference between revisions)
 Revision as of 16:19, 20 October 2013 (view source) (→October 19, 2013)← Previous diff Revision as of 16:20, 20 October 2013 (view source) (→October 19, 2013)Next diff → Line 22: Line 22: |} |} - ''Calculations'' + ''Calculations'' - ng insert = bp insert/bp vector * 2 * 50
+ ng insert = bp insert/bp vector * 2 * 50
ng insert = 588/4611 * 2 *50 = 12.75211 ng ng insert = 588/4611 * 2 *50 = 12.75211 ng - μL insert = ng insert/concentration insert
+ μL insert = ng insert/concentration insert
μL insert = 12.75211/15.535 = .82μL μL insert = 12.75211/15.535 = .82μL - μL vector = 50 ng/concentration vector
+ μL vector = 50 ng/concentration vector
μL vector = 50/20.838 = 2.4  μL μL vector = 50/20.838 = 2.4  μL

## Revision as of 16:20, 20 October 2013

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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
ng insert = 588/4611 * 2 *50 = 12.75211 ng

μL insert = ng insert/concentration insert
μL insert = 12.75211/15.535 = .82μL

μL vector = 50 ng/concentration vector
μL vector = 50/20.838 = 2.4 μL

 1 Control DNA Insert 0.8 0 DNA Vector (MV9) 2.4 2.4 T4 Ligase 1 1 Lign Buffer (2x) 5 5 dH2O 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 + dH2O.
• 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.