Hypothesis 2: Gene L is necessary for phage propagation.
- Unfortunately, gel electrophoresis indicated both primer 4 and primer 4 T3485A samples did not work.
- Here is a list of things that changed from yesterday until today:
- new dNTPs mix: shouldn't be a problem, so will continue to use this mix.
- re-programmed PCR program: I checked the machine. The program is as I have it here, so it should work, since the same (or similar) cycling parameters worked previously.
- different primer mixes: this I'm not so sure of. I will create new mixes at 100 μM and 10 μM and try again.
Characterization B-C: Expression of PHIX174 promoters/UTRs fused to PX-UTR1-deGFP and PX-UTRX-deGFP.
- Gel electrophoresis indicated BamXI/XhoI double digested DNA of:
- pBEST-PA-MGapt-T500 ~ 2500 bp dsDNA - OK
- pBEST-PB-MGapt-T500 ~ 2500 bp dsDNA - OK
- pBEST-PD-MGapt-T500 ~ 2500 bp dsDNA - OK
- pBEST-PF-MGapt-T500 ~ 3000-3500 bp dsDNA - not OK
- pBEST-PG-MGapt-T500 ~ 2500 bp dsDNA - OK
- pBEST-PL-MGapt-T500 ~ 2500 bp dsDNA - OK
- pBEST-PL-PA-MGapt-T500 ~ smear - not OK
- pBEST-NONE-MGapt-T500 ~ 2500 bp dsDNA - OK
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PHIX174 Cell Free Expression
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