User:Sean P Corum/Notebook/PHIX174 Cell Free/2012/06/29: Difference between revisions
From OpenWetWare
Sean P Corum (talk | contribs) mNo edit summary |
Sean P Corum (talk | contribs) |
||
Line 9: | Line 9: | ||
* I peformed hybridization on oligonucleotides to construct the following: | * I peformed hybridization on oligonucleotides to construct the following: | ||
** SphI-PA | ** SphI-PA-NcoI linker at 100 nM final concentration | ||
** SphI-PD | ** SphI-PD-NcoI linker at 100 nM final concentration | ||
** SphI-PF | ** SphI-PF-NcoI linker at 100 nM final concentration | ||
* I peformed [http://openwetware.org/index.php?title=Corum:PCR PCR] followed by [http://openwetware.org/index.php?title=Corum:PCR_Purification PCR purification], speedvac down to ~20 μL, digestion with SphI and NcoI, and another round of [http://openwetware.org/index.php?title=Corum:PCR_Purification PCR purification] to create the following linkers: | * I peformed [http://openwetware.org/index.php?title=Corum:PCR PCR] followed by [http://openwetware.org/index.php?title=Corum:PCR_Purification PCR purification], speedvac down to ~20 μL, digestion with SphI and NcoI, and another round of [http://openwetware.org/index.php?title=Corum:PCR_Purification PCR purification] to create the following linkers: | ||
** SphI-PB | ** SphI-PB-NcoI linker | ||
** SphI-PG | ** SphI-PG-NcoI linker | ||
** SphI-PL | ** SphI-PL-NcoI linker | ||
** SphI-PLPA | ** SphI-PLPA-NcoI linker | ||
* All of the linkers should be at ~100 nM final concentration, so they can be used with the pBEST-SphI//NcoI-deGFP-T500 linker (10 nM) in ligation to create the desired set of pBEST-PX-UTR1-deGFP-T500 constructs. | * All of the linkers should be at ~100 nM final concentration, so they can be used with the pBEST-SphI//NcoI-deGFP-T500 linker (10 nM) in ligation to create the desired set of pBEST-PX-UTR1-deGFP-T500 constructs. | ||
Revision as of 11:24, 29 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> </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.
Characterization C: Expression of PHIX174 promoters fused to UTRX-deGFP.
Hypothesis 2: Gene L is necessary for phage propagation.
To do
NotesRecently Edited Notebook Pages Digital Signature
|