Wisconsin-Madison/11 July 2009

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