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Team:Queens-Canada/30 August 2010
From 2010.igem.org
August 30, 2010
Liquid Culture
Thai
pOsm-10::eCFP::unc-54 3'UTR::1A3, pSra-10::eCFP::unc-54 3'UTR::1A3
Digestion
Hao and Thai
pSip-1::1C3, pFlp-1::1C3, pOdr-10::1C3, mCherry::1C3
| EcoRI and SpeI | XbaI and PstI | EcoRI and PstI |
| pStr-220::1C3 | eCFP::unc-54 3'UTR::1T3 | pSB1T3 |
| mCherry::1C3 | unc-54 3'UTR::1C3 | |
| pFlp-1::1C3 | ||
| pSip-1::1C3 | ||
| pOdr-10::1C3 |
Backbone treated with CIAP for 30 mins.
Ligation
Hao and Thai
pSip-1, pStr-220, pFlp-1, pOdr-10::eCFP::unc-54 3'UTR::1A3, ChR2::unc-54 3'UTR::1T3
| EcoRI and SpeI | XbaI and PstI | EcoRI and PstI |
| pStr-1 | eCFP::unc-54 3'UTR | 1A3 |
| pStr-220 | eCFP::unc-54 3'UTR | 1A3 |
| pFlp-1 | eCFP::unc-54 3'UTR | 1A3 |
| pOdr-10 | eCFP::unc-54 3'UTR | 1A3 |
| mCherry | unc-54 3'UTR | 1T3 |
Backbone treated with CIAP for 30 mins.
Heat Shock
Hao
pSip-1, pStr-220, pFlp-1, pOdr-10::eCFP::unc-54 3'UTR::1A3, ChR2::unc-54 3'UTR::1T3
Transformed ligation products from 30/08/10
Microinjection
Hao
pMec-7::eCFP::unc-54 3'UTR::1C3
Transferred F1 transgenic worms to new plate
