Revision as of 14:43, 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

Transformation into Top10 E. coli

pSB1C3/IRES
pSB1C3/IaaM
pSB1C3/IaaH
pSB1C3/IaaH-IRES
pSC2+/IaaH
pCS2+/IRES

@@ Line 27: / Line 27: @@
 <h3>Transformation into <i>Top10 E. coli</i> </h3>
+pSB1C3/IRES<br>
+pSB1C3/IaaM<br>
+pSB1C3/IaaH<br>
+pSB1C3/IaaH-IRES<br>
+pSC2+/IaaH<br>
+pCS2+/IRES<br>
 </html>

Team:Evry/Notebook/w10

From 2012.igem.org