Wisconsin-Madison/11 July 2009
From 2009.igem.org
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July 11, 2009
Ex 18: pTaq
De-stained protein gel
Ex 20: New Cloning Methods
Make Electro Competent Cells
Double Digestion of BB (E+S)
1. YhfR (560+2157=) 2717 bp
2. NudF (560+2157=) 2717 bp
3. ProU (300+2157=) 2457 bp
4. Choline (2000+2157=) 4157 bp
5. BB (2157+1069=) 3226 bp control
18 uL RXN:
1. 11uL H2O
2. 1.8uL Buffer 3
3. 0.18uL Protein BSA
4. 3uL DNA
5. 1ul Enzyme EcoR1
6. 1uL Enzyme Spe1
7. (2hr waterbath) 3.6 uL Dye
Undigested(NudF2, ProU, Cho, BB, YhfR1, YhfR2, NudF1), Ladder, Digested(ProU, Cho, BB+Red, YhfR1, YhfR2, NudF1, NudF2)
Results: We have YhfR as our only BioBrick
Why do we not have two bands for ProU, Cho, and NudF 1 and 2
Writing Project Description
Writing Paper for Project
Shotgun Cloning
- added 100mL media A to 7/8 natural transformation test cultures and 100uL CM - NP