Team:Queens-Canada/3 August 2010

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August 3, 2010

Liquid Culture

Steve and Chris

unc-54 3'UTR::1A3, pRab-7::1A3, pMec-7::1A3

Digestion-Ligation

Thai and Hao

mCherry, pCeh-12, pSrb-6, pOdr-1, pOdr-10, NpHR, eYFP, pHsp-3, pStr-220, pFlp-1, pOsm-10, pStr-1, eCFP, pSB1C3

Cut with EcoRI and PstI into pSB1C3. Backbone treated with CIAP. Used T4 ligase.

Heat Shock

mCherry, pCeh-12, pSrb-6, pOdr-1, pOdr-10, NpHR, eYFP, pHsp-3, pStr-220, pFlp-1, pOsm-10, pStr-1, eCFP, pSB1C3

Transformed TOP10 using previous ligation products.

PCR

Chris

pFlp-1, pHsp-3, NpHR

Miniprep

Chris and Mary and Louis

Solution II used in place of Solution I.

Microinjection

Hao

Practiced injections with Sur-5::GFP marker.