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

From OpenWetWare
Jump to navigationJump to search
Line 8: Line 8:
===Characterization B: Expression of PHIX174 promoters fused to UTR1-deGFP.===
===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:
* Performed [http://openwetware.org/wiki/Corum:T4_Ligation 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-PB-UTR1-deGFP-T500
*# pBEST-PD-UTR1-deGFP-T500
*# pBEST-PD-UTR1-deGFP-T500

Revision as of 18:25, 10 July 2012

PHIX174 Cell Free Expression <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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.
Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Sean_P_Corum/Notebook/PHIX174_Cell_Free/2012/07/11&oldid=613478"