Team:Evry/Notebook/w9

From 2012.igem.org

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Issue: Not enough DNA for ligation.
Issue: Not enough DNA for ligation.
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Mini-prep pCS2+-dendra2 and pCS2+-mcherry checked by gel.
+
Mini-prep pCS2+-dendra2 and pCS2+-mcherry checked by gel.<br>
-
inolation for midi-prep pNHK60 (Tir1/GFP-aid),
+
inolation for midi-prep pNHK60 (Tir1/GFP-aid),<br>
-
inoculation for mini-prep pSB1C3 K515100 (IaaH+IaaM).
+
inoculation for mini-prep pSB1C3 K515100 (IaaH+IaaM).<br>
<h2>Tuesday, 7th August</h2>
<h2>Tuesday, 7th August</h2>
</br>
</br>
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Midi-prep pNHK60 (Tir1/GFP-aid),
+
Midi-prep pNHK60 (Tir1/GFP-aid),<br>
-
Mini-prep pSB1C3 K515100 (IaaH+IaaM).
+
Mini-prep pSB1C3 K515100 (IaaH+IaaM).<br>
-
Digestion pSB1C3 K515100 (IaaH+IaaM)with EcoRI and PstI then transformation in pCS2+ (amplified with PCR, digested with EcoRI and PstI then purified).
+
Digestion pSB1C3 K515100 (IaaH+IaaM)with EcoRI and PstI then transformation in pCS2+ (amplified with PCR, digested with EcoRI and PstI then purified).<br>
-
 
+
 +
<h2>Wednesday, 8th August</h2>
 +
<h3>Iaa BB</h3>
 +
IaaH PCR: P35 + P33 + pSBIC3 IaaH IaaM K515100 <br>
 +
IaaM PCR: P34 + P22 + pSBIC3 IaaH IaaM K515100<br>
 +
IRES PCR: P31 + P32 + pNHK60<br>
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</html>

Revision as of 10:06, 8 August 2012

Weeks:

June July August September October November

Week 9: 6th August - 12th August

Monday, 6th August

Reporter test

We had a lot of reporters BB but we didn't know if they're all working. We test them with a electrophoresis: Two RFP, a GFP, a CFP and a YFP worked.

GFP BB

Digestion of GFP BB with EcoRI and PstI, gel extraction and DNA purification. Goal: insertion in pCS2+ BB Issue: Not enough DNA for ligation. Mini-prep pCS2+-dendra2 and pCS2+-mcherry checked by gel.
inolation for midi-prep pNHK60 (Tir1/GFP-aid),
inoculation for mini-prep pSB1C3 K515100 (IaaH+IaaM).

Tuesday, 7th August


Midi-prep pNHK60 (Tir1/GFP-aid),
Mini-prep pSB1C3 K515100 (IaaH+IaaM).
Digestion pSB1C3 K515100 (IaaH+IaaM)with EcoRI and PstI then transformation in pCS2+ (amplified with PCR, digested with EcoRI and PstI then purified).

Wednesday, 8th August

Iaa BB

IaaH PCR: P35 + P33 + pSBIC3 IaaH IaaM K515100
IaaM PCR: P34 + P22 + pSBIC3 IaaH IaaM K515100
IRES PCR: P31 + P32 + pNHK60