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

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(DD I14032.B0034 Mixture)
Line 12: Line 12:
==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 19:30, 16 August 2007

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

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