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Team:Evry/Notebook/w12
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| - | <h1> | + | <h1>Week 12: 27th August - 2nd September</h1> |
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| + | <h2>Monday 27th</h2> | ||
| + | |||
| + | Preparation of different cloning: <br> | ||
| + | * Amplification of IAAH and IAAM with new primers <br> | ||
| + | * Amplification of citrine, sfGFP and mCFP with kozac sequence <br> | ||
| + | * Digestion of GFP-AID, BOO34 GB, E1010 GB, R0010 GB, B0015 GB, GB insert in EcoRI PstI <br> | ||
| + | |||
| + | New cloning attempt of IAAH and IAAM in pSC2+ | ||
| + | |||
| + | <h2>Tuesday 28th</h2> | ||
| + | |||
| + | *Preparation of digested pSB1C3 in high quantity. Digestion of pSC2+ <br> | ||
| + | *Digestion of the IAAH-IAAM, citrine, sfGFP, mCFP <br> | ||
| + | |||
| + | Ligation of all the parts digested the day before and the day. Transformation using the new electrocompetent cell protocol. Concentration before plating and incubation overnight. <br> | ||
| + | |||
| + | Ligation of TirI using the in-fusion protocol and electroporation <br> | ||
| + | |||
| + | New transformation of pSB1C3 from plates <br> | ||
| + | |||
| + | New culture of pSC2+ BB for midi-prep the day after <br> | ||
| + | |||
| + | <h2>Wednesday 29th</h2> | ||
| + | |||
| + | All the plates contained cloned. The color of the GB inset indicates that the insertion might have been efficient for most of the clonings. <br> | ||
| + | |||
| + | Colony PCR of 8 clones of each constructs carried out. | ||
<table> | <table> | ||
| - | <tr> | + | <tr><td>GFP-AID</td><td>pSC2+</td><td>clones XXXXX</td></tr> |
| - | + | <tr><td>citrine</td><td>pSC2+</td><td>clones XXXXX</td></tr> | |
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| - | </tr> | + | |
</table> | </table> | ||
| - | < | + | <td><tr>GG insert</tr><tr>pSC2+</tr><tr>clones XXXXX</tr></td> |
| + | <td><tr>sfGFP</tr><tr>pSC2+</tr><tr>clones XXXXX</tr></td> | ||
| + | <td><tr>mCFP</tr><tr>pSC2+</tr><tr>clones XXXXX</tr></td> | ||
| + | <td><tr>GG insert</tr><tr>pSB1C3</tr><tr>clones XXXXX</tr></td> | ||
| + | <td><tr>E1010</tr><tr>pSB1C3</tr><tr>clones XXXXX</tr></td> | ||
| + | <td><tr>B0034</tr><tr>pSB1C3</tr><tr>clones XXXXX</tr></td> | ||
| + | <td><tr>B0015</tr><tr>pSB1C3</tr><tr>clones XXXXX</tr></td> | ||
| + | <td><tr>R1010</tr><tr>pSB1C3</tr><tr>clones XXXXX</tr></td> | ||
| + | <td><tr>GFP-AID</tr><tr>pSB1C3</tr><tr>clones XXXXX</tr></td> | ||
| + | <td><tr>TirI</tr><tr>pSB1C3</tr><tr>clones XXXXX</tr></td> | ||
</html> | </html> | ||
Latest revision as of 11:45, 1 September 2012
Weeks:
| June | July | August | September | October | November | ||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Week 12: 27th August - 2nd September
Monday 27th
Preparation of different cloning:* Amplification of IAAH and IAAM with new primers
* Amplification of citrine, sfGFP and mCFP with kozac sequence
* Digestion of GFP-AID, BOO34 GB, E1010 GB, R0010 GB, B0015 GB, GB insert in EcoRI PstI
New cloning attempt of IAAH and IAAM in pSC2+
Tuesday 28th
*Preparation of digested pSB1C3 in high quantity. Digestion of pSC2+*Digestion of the IAAH-IAAM, citrine, sfGFP, mCFP
Ligation of all the parts digested the day before and the day. Transformation using the new electrocompetent cell protocol. Concentration before plating and incubation overnight.
Ligation of TirI using the in-fusion protocol and electroporation
New transformation of pSB1C3 from plates
New culture of pSC2+ BB for midi-prep the day after
Wednesday 29th
All the plates contained cloned. The color of the GB inset indicates that the insertion might have been efficient for most of the clonings.Colony PCR of 8 clones of each constructs carried out.
| GFP-AID | pSC2+ | clones XXXXX |
| citrine | pSC2+ | clones XXXXX |
