IGEM:PKU Beijing/2009/Notebook/Bistable/2009/07/19/Shan Shen

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10:30 Miniprep 2-4O, 1-18P, low-copy bi-stable system Concentration:

Number of the plasmids Concentration(ng/μL)
2-4O 350
1-18P 250
Low-copy bi-stable system 125

12:00 Digest 1-18P, 2-4O, low-copy plasmids. 1-18P by Spe1 and Pst1; 2-4O by Xba1 and Pst1; low-copy bi-stable system by EcoR1 and Pst1.

18:00 Recycle those digestion products.

22:40 Ligation of high-copy bi-stable system fragments and high-copy backbone.

Total 10μL
Vectors 1μL
Inserts 7μL
T4 ligase 1μL
Buffer 1μL