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

From OpenWetWare

Jump to: navigation, search

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.

Total20μL
Plasmids4μL
EcoR11μL
Pst11μL
Buffer2μL
ddH2O12μL

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:

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 plasmidsConcentration(ng/μL)
1-18A250
1-18C350

23:00
Digest 1-18A.

Total20μL
Plasmids4μL
Spe11μL
Pst11μL
Buffer2μL
ddH2O12μL
Personal tools