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Team:Evry/Notebook/w9
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<h2>Tuesday, 7th August</h2> | <h2>Tuesday, 7th August</h2> | ||
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| - | Midi-prep pNHK60 (Tir1/GFP-aid),<br> | + | Midi-prep pNHK60 (Tir1/GFP-aid): elution in 1ml, 35,2ng/ul<br> |
Mini-prep pSB1C3 K515100 (IaaH+IaaM).<br> | 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).<br> | Digestion pSB1C3 K515100 (IaaH+IaaM)with EcoRI and PstI then transformation in pCS2+ (amplified with PCR, digested with EcoRI and PstI then purified).<br> | ||
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<h2>Wednesday, 8th August</h2> | <h2>Wednesday, 8th August</h2> | ||
| + | Speed vac of pNHK60 (midi-prep 7th August): resuspension in 50ul : 1254,9ng/ul, 260/280nm = 1,79 (tube A9)<br> | ||
| + | inolation for midi-prep pSB1C3 (IaaH+IaaM),<br> | ||
<h3>Iaa BB</h3> | <h3>Iaa BB</h3> | ||
IaaH PCR: P35 + P33 + pSBIC3 IaaH IaaM K515100 <br> | IaaH PCR: P35 + P33 + pSBIC3 IaaH IaaM K515100 <br> | ||
IaaM PCR: P34 + P22 + pSBIC3 IaaH IaaM K515100<br> | IaaM PCR: P34 + P22 + pSBIC3 IaaH IaaM K515100<br> | ||
IRES PCR: P31 + P32 + pNHK60<br> | IRES PCR: P31 + P32 + pNHK60<br> | ||
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| + | <h2>Thursday, 9th August</h2> | ||
| + | Midi-prep pSB1C3 (IaaH+IaaM)<br> | ||
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</html> | </html> | ||
Revision as of 09:56, 9 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): elution in 1ml, 35,2ng/ulMini-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
Speed vac of pNHK60 (midi-prep 7th August): resuspension in 50ul : 1254,9ng/ul, 260/280nm = 1,79 (tube A9)inolation for midi-prep pSB1C3 (IaaH+IaaM),
Iaa BB
IaaH PCR: P35 + P33 + pSBIC3 IaaH IaaM K515100IaaM PCR: P34 + P22 + pSBIC3 IaaH IaaM K515100
IRES PCR: P31 + P32 + pNHK60
