User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/06/22: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 13: Line 13:
===Characterization B: Expression of PHIX174 promoters fused to UTR1-deGFP.===
===Characterization B: Expression of PHIX174 promoters fused to UTR1-deGFP.===


* Performed miniprep plasmid isolation on the five clones.
* Performed [http://openwetware.org/wiki/Corum:miniprep miniprep plasmid isolation] on the five clones.
* Submitted for sequencing.
* Submitted samples for [http://openwetware.org/wiki/Corum:DNA_Sequencing sequencing].


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

Revision as of 13:36, 25 June 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>


June 22, 2012

  • SC 15:43, 23 June 2012 (EDT):

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

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

  • Yesterday, I digested PCR products with SphI and NcoI to create SphI-(PB-UTRB, PG-UTRG, PL-UTRL, and PL-PA-UTRA)-NcoI linkers. Today, I performed gel extraction on the digest products.
  • I also hybridized oligonucleotides to create SphI-(PA-UTRA, PD-UTRD, PF-UTRF)-NcoI linkers (100μM, 10μM, 1μM, and 100nM serial dilutions).
  • The next step will be to ligate, isolation plasmid clones, and sequence.

Hypothesis 2: Gene L is necessary for phage propagation.

  • Need to design new primers (regular and mutagenic) for amplifying PHIX174 genome.


Notes

  • None.


Recently Edited Notebook Pages


Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Sean_P_Corum/Notebook/PHIX174_Cell_Free/2012/06/22&oldid=610031"