IGEM:MIT/2007/Notebook/2007-8-16

Agenda

1. Make electro/chemically competent cells
2. Transform and plate R0051
3. Make more DD R0051
4. Redo R0051.B0034 ligation
5. DD I14032.B0034 and put it into pSB1AC3 as a blank vector
6. Run Polystyrene binding efficiency assay

Making Electrochemically competent cells

Using BL-21 (for higher protein expression) Followed TK's OWW protocol.

DD I14032.B0034 Mixture

For blank (control) vector:
5 µL I14032.B0034 #4 (225.7 ng/µL)

1 µL EcoRI

1 µL PstI

0.5 µL BSA

5 µL NEB Buffer 3

37.5 µL H20

For 3A assembly with CPX:
5 µL I14032.B0034 #4 (225.7 ng/µL)

1 µL EcoRI

1 µL SpeI

0.5 µL BSA

5 µL NEB Buffer 2

37.5 µL H20

put into 37C at 12:15 PM