User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/09/21

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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:
    1. pBEST-PA-MGapt-T500 ~ 2500 bp dsDNA - OK
    2. pBEST-PB-MGapt-T500 ~ 2500 bp dsDNA - OK
    3. pBEST-PD-MGapt-T500 ~ 2500 bp dsDNA - OK
    4. pBEST-PF-MGapt-T500 ~ 3000-3500 bp dsDNA - not OK
    5. pBEST-PG-MGapt-T500 ~ 2500 bp dsDNA - OK
    6. pBEST-PL-MGapt-T500 ~ 2500 bp dsDNA - OK
    7. pBEST-PL-PA-MGapt-T500 ~ smear - not OK
    8. pBEST-NONE-MGapt-T500 ~ 2500 bp dsDNA - OK