User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/06/25

PHIX174 Cell Free Expression

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Characterization B: Expression of PHIX174 promoters fused to UTR1-deGFP. Waiting for sequencing results. Characterization C: Expression of PHIX174 promoters fused to UTRX-deGFP. Isolated pBEST-SphI//NcoI-deGFP-T500 vector backbone digest product via gel extraction. Gel eletrophoresis observation: expected 3096 bp and 106 bp dsDNA bands, observed . The larger band was the one purified from the gel. I ligated the different linkers into the pBEST-SphI//NcoI-deGFP-T500 backbone to form pBEST-PX-UTRX-deGGFP-T500, where X = A, B, D, F, G, L, and LA. Hypothesis 2: Gene L is necessary for phage propagation. Need to design new primers (regular and mutagenic) for amplifying PHIX174 genome.

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SC 16:45, 25 June 2012 (EDT):