Revision as of 10:31, 13 August 2012

Weeks:

June			July				August				September				October				November

Week 10: 13th August - 19th August

Monday, 13th August

Tadpoles injection (pNHK60)

Preparation of pNHK60 dilution(phenol/chloro extraction of Friday, 10th August)
Tube A12 (50ng/ul): 1,695ul A11 + 73,305ul H2Odd
Tube A13 (25ng/ul): 25ul A12 + 25ul H2Odd

PCR

TirI

Primers: P1 and P2
ADN: TirI-PCR P2-P36 (08.08) + TirI PCR P1-P4 (10.08)

pCS2+

Primers: P7+P8
ADN:A1

Cloning pSB1C3/IRES, pSB1C3/IaaM, pSB1C3/IaaH, pSC2+/IaaH (to biobrick pCS2+)

Restriction with EcoRI and PstI for pSB1C3 (K515100) then gel extraction of the backbone
Restriction with EcoRI and PstI for product of PCR: IRES, IaaH, IaaM and pCS2+ then purification with PCR clean up kit
Then ligation of pCS2+ with IaaH, pSB1C3 with IRES, pSB1C3 with IaaM and pSB1C3 with IaaH

Injection of PNHK60 into Xenopus eggs

Directly after fertilization of xenopus tropicalis eggs: injection of pNHK60 (plasmid with Tir1 and GFP-aid)
The production of GFP is expected into eggs

Retrieved from "http://2012.igem.org/Team:Evry/Notebook/w10"

@@ Line 16: / Line 16: @@
 Primers: P7+P8<br>
 ADN:A1<br>
+<h3>Cloning pSB1C3/IRES, pSB1C3/IaaM, pSB1C3/IaaH, pSC2+/IaaH (to biobrick pCS2+) </h3>
+Restriction with EcoRI and PstI for pSB1C3 (K515100) then gel extraction of the backbone<br>
+Restriction with EcoRI and PstI for product of PCR: IRES, IaaH, IaaM and pCS2+ then purification with PCR clean up kit<br>
+Then ligation of pCS2+ with IaaH, pSB1C3 with IRES, pSB1C3 with IaaM and pSB1C3 with IaaH<br>
+<h3>Injection of PNHK60 into Xenopus eggs<h3>
+Directly after fertilization of xenopus tropicalis eggs: injection of pNHK60 (plasmid with Tir1 and GFP-aid)<br>
+The production of GFP is expected into eggs<br>
 </html>

Team:Evry/Notebook/w10

From 2012.igem.org