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From 2012.igem.org

Weeks:

June			July				August				September				October				November

Week 11: 20th August - 26th August

Monday, 20th August

pSC2+ RFP, pSC2+ IaaH

Transformation results: 2 colonies pink on pSC2+ RFP plate, no one on the pSC2+ IaaH plate PCR colony on the 2 pink colonies pSC2+ RFP with P26 and P27 primers

IaaH-IaaM-IRES

After the PCR of IaaH, IaaM and IRES (16th August) digestion with EcoRI and PstI (but also or pSB1C3) then gel extraction
Ligation of Ires, IaaH and IaaM in pSB1C3

Tuesday, 21st August

Transformation of pSB1C3-IRES, pSB1C3-IaaH and pSB1C3-IaaM plus control ligation pSB1C3 without insert
Injection pNHK60 (50ng/microL) into xenopus tropicalis eggs

Wednesday, 22nd August

Embryos after In Vitro fertilization

1- day old embryos were sorted under microscope (live/dead), counted and moved to 12-well plate.
Auxin intake experiment:
10 embryos placed in each well, then MMR medium added with different concentration of auxin (0µM, 125µM, 250µM, 500µM)
Incubation in 21°C

Thursday, 23rd August

Metabolites extraction from embryos

Metabolites were extracted from 2-days old embryos as described in Protocols (preparation of HPLC test)

Friday, 24th August

Metabolites extraction from embryos

Metabolites were extracted from 3-days old embryos as described in Protocols (preparation of HPLC test)