User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/07/11: Difference between revisions

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(Autocreate 2012/07/11 Entry for User:Sean_P_Corum/Notebook/PHIX174_Cell_Free)
 
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==Entry title==
===Characterization B: Expression of PHIX174 promoters fused to UTR1-deGFP.===
* Insert content here...


* Performed ligation of latest batch of SphI-PX-NheI linkers and pBEST-SphI//NheI-UTR1-deGFP-T500 backbone to obtain the following products:
*# pBEST-PB-UTR1-deGFP-T500
*# pBEST-PD-UTR1-deGFP-T500
*# pBEST-PF-UTR1-deGFP-T500
*# pBEST-PG-UTR1-deGFP-T500
*# pBEST-PL-UTR1-deGFP-T500
*# pBEST-PLPA-UTR1-deGFP-T500
*# -linker control


===Characterization C: Expression of PHIX174 promoters fused to UTRX-deGFP.===
* I transformed pBEST-PA-UTRA-deGFP-T500 ligation product into JM109. Plates showed many colonies. -linker negative control showed fewer colonies, as expected. Six colonies were picked from the plate for miniculture.
* Performed ligation of latest batch of SphI-PX-UTRX-NcoI linkers and pBEST-SphI//NheI-UTR1-deGFP-T500 backbone to obtain the following products:
*# pBEST-PB-UTRB-deGFP-T500
*# pBEST-PD-UTRD-deGFP-T500
*# pBEST-PF-UTRF-deGFP-T500
*# pBEST-PG-UTRG-deGFP-T500
*# pBEST-PL-UTRL-deGFP-T500
*# -linker control
===Hypothesis 2: Gene L is necessary for phage propagation.===
* On hold.
   
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Revision as of 18:24, 10 July 2012

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Characterization B: Expression of PHIX174 promoters fused to UTR1-deGFP.

  • Performed ligation of latest batch of SphI-PX-NheI linkers and pBEST-SphI//NheI-UTR1-deGFP-T500 backbone to obtain the following products:
    1. pBEST-PB-UTR1-deGFP-T500
    2. pBEST-PD-UTR1-deGFP-T500
    3. pBEST-PF-UTR1-deGFP-T500
    4. pBEST-PG-UTR1-deGFP-T500
    5. pBEST-PL-UTR1-deGFP-T500
    6. pBEST-PLPA-UTR1-deGFP-T500
    7. -linker control

Characterization C: Expression of PHIX174 promoters fused to UTRX-deGFP.

  • I transformed pBEST-PA-UTRA-deGFP-T500 ligation product into JM109. Plates showed many colonies. -linker negative control showed fewer colonies, as expected. Six colonies were picked from the plate for miniculture.
  • Performed ligation of latest batch of SphI-PX-UTRX-NcoI linkers and pBEST-SphI//NheI-UTR1-deGFP-T500 backbone to obtain the following products:
    1. pBEST-PB-UTRB-deGFP-T500
    2. pBEST-PD-UTRD-deGFP-T500
    3. pBEST-PF-UTRF-deGFP-T500
    4. pBEST-PG-UTRG-deGFP-T500
    5. pBEST-PL-UTRL-deGFP-T500
    6. -linker control

Hypothesis 2: Gene L is necessary for phage propagation.

  • On hold.
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