IGEM:PKU Beijing/2009/Notebook/Bistable/2009/07/12/Shan Shen

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Prepare competent cells.
Construction

8:00
The order of the samples: marker, plasmid control of 1-18A, digestion product of 1-18A
The digestion products run faster than the plasmids.

9:30
Shake 1-18A and 1-18C again.

15:30
Electrophoresis to test the digestion products
The order and the amount of the samples: marker 5μL, plasmids of high-copy bi-stable system 3μL, digestion products of hi-copy bi-stable system 20μL, plasmids of low-copy bi-stable system 5μL, digestion products of low-copy bi-stable system 20μL.
Results: Bands are smeared.

16:20
Digest high-copy bi-stable system and low-copy bi-stable system again.

Total 20μL
Plasmids 4μL
EcoR1 1μL
Pst1 1μL
Buffer 2μL
ddH2O 12μL

22:00
Electrophoresis to test the digestion products
The order and the amount of the samples: marker 5μL, plasmids of high-copy bi-stable system 3μL, digestion products of hi-copy bi-stable system 20μL digestion products of low-copy bi-stable system 20μL.
Results:
PKU 20090712 Shan Shen 1.JPG
High- copy plasmids have been successfully digested, while the low-copy plasmids are still digested to be smeared.

22:30
Miniprep of 1-18A and 1-18C

Number of the plasmids Concentration(ng/μL)
1-18A 250
1-18C 350

23:00
Digest 1-18A.

Total 20μL
Plasmids 4μL
Spe1 1μL
Pst1 1μL
Buffer 2μL
ddH2O 12μL