EPF-Lausanne/25 August 2009

From 2009.igem.org

(Difference between revisions)
(→Trp Operon synthesis)
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|-
| width="33%" |
| width="33%" |
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[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]
+
dNTP
| width="33%" align="center"|
| width="33%" align="center"|
-
Promoter LacI
+
1ul each
| width="34%" align="center" |
| width="34%" align="center" |
-
A
+
1ul each
-
| width="50%" align="center" |
+
-
1D (1)
+
|-
|-
| width="33%" |
| width="33%" |
-
[http://partsregistry.org/Part:BBa_B0030 BBa_B0030]
+
template primers
| width="33%" align="center"|
| width="33%" align="center"|
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RBS
+
1ul (RV) + 1.5ul (FW) each
| width="34%" align="center" |
| width="34%" align="center" |
-
A
+
-
-
| width="50%" align="center" |
+
-
1H (1)
+
|-
|-
| width="33%" |
| width="33%" |
-
[http://partsregistry.org/Part:BBa_E0240 BBa_E0240]
+
Klenow enzyme
| width="33%" align="center"|
| width="33%" align="center"|
-
RBS-GFP-TER
+
1 ul each
| width="34%" align="center" |
| width="34%" align="center" |
-
A
+
1 ul
-
| width="50%" align="center" |
+
-
12M (1)
+
|-
|-
| width="33%" |
| width="33%" |
-
[http://partsregistry.org/Part:BBa_I13507 BBa_I13507]
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dH2O
| width="33%" align="center"|
| width="33%" align="center"|
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RBS-mRFP-TER
+
18.5 ul each
| width="34%" align="center" |
| width="34%" align="center" |
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A
+
21ul
-
| width="50%" align="center" |
+
|----------------------------
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22O (1)
+
-
|-
+
| width="33%" |
| width="33%" |
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[http://partsregistry.org/Part:BBa_J13002 BBa_J13002]
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total
| width="33%" align="center"|
| width="33%" align="center"|
-
pTetR-RBS
+
25 ul each
| width="34%" align="center" |
| width="34%" align="center" |
-
A
+
25 ul
-
| width="50%" align="center" |
+
-
13B (1)
+
|-
|-
|}
|}

Revision as of 11:56, 2 September 2009



Contents

25 August 2009




Wet Lab

Miniprep:

Made according to the protocol of Aisgen Miniprep Kit Elution in 50 ul. The RD2 #5 plasmids prep as some that have already been prepared will be sent for sequencing this afternoon.


Mediums

New medium were made to grow the double-transformed DH5 a (LovTap BB and RO2) -> should be without thy.

M9 was redone, pH adjusted with amino acid and filtred. for 1L:

5g glucose
6g Na2PO4
3g KH2PO4
1g NH4Cl
0.5g NaCl
0.12g MgSO4
0.01g CaCl2


Trp Operon synthesis

Using the Klenow fragments protocol and primers:£

Trp operon Rev
Trp operon Fwd
Parts&Characteristics
2x Trp 1x Negative Control

NEB 2 (10x)

2ul each

2ul

dNTP

1ul each

1ul each

template primers

1ul (RV) + 1.5ul (FW) each

-

Klenow enzyme

1 ul each

1 ul

dH2O

18.5 ul each

21ul

total

25 ul each

25 ul

People in the lab

Nath, Basile, Gab, Christian



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