IGEM:MIT/2007/Notebook/2007-8-16: Difference between revisions
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==DD I14032.B0034 Mixture== | ==DD I14032.B0034 Mixture== | ||
For blank (control) vector: | |||
<br> | |||
5 µL I14032.B0034 #4 (225.7 ng/µL) | 5 µL I14032.B0034 #4 (225.7 ng/µL) | ||
1 µL EcoRI | 1 µL EcoRI | ||
1 µL PstI | 1 µL PstI | ||
0.5 µL BSA | 0.5 µL BSA | ||
5 µL NEB Buffer 3 | 5 µL NEB Buffer 3 | ||
37.5 µL H20 | |||
For 3A assembly with CPX: | |||
<br> | |||
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 | 37.5 µL H20 | ||
put into 37C at 12:15 PM | put into 37C at 12:15 PM | ||
Revision as of 16:30, 16 August 2007
Agenda
- Make electro/chemically competent cells
- Transform and plate R0051
- Make more DD R0051
- Redo R0051.B0034 ligation
- DD I14032.B0034 and put it into pSB1AC3 as a blank vector
- 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
