Team:Evry/Notebook/w9
From 2012.igem.org
(Difference between revisions)
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Issue: Not enough DNA for ligation. | Issue: Not enough DNA for ligation. | ||
- | 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> | ||
- | 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> | ||
</html> | </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 K515100IaaM PCR: P34 + P22 + pSBIC3 IaaH IaaM K515100
IRES PCR: P31 + P32 + pNHK60