Team:Hawaii/Notebook/2008-09-24

From 2008.igem.org

(Difference between revisions)
(Making Emergency BB Vector Insert (contaminant troubleshoot))
 
Line 28: Line 28:
::* PCR machine froze while ExoSAPing. Unable to retrieve samples from machine. Jamie says she will figure it out.
::* PCR machine froze while ExoSAPing. Unable to retrieve samples from machine. Jamie says she will figure it out.
-
===Making Emergency BB Vector Insert (contaminant troubleshoot)===
+
===Making Emergency BB Vector Insert (help contaminant troubleshoot)===
[[Image:20080924-emergency_vector_pRLBB_vector_insert_PCR_redo.annotated.jpg|thumb|right|making x6 Parallel MCS Replacement fragmenets via PCR Redo!!! with clean uncontaminated primers [[Media:20080924-emergency_vector_pRLBB_vector_insert_PCR_redo.jpg|download unannotated]]]]
[[Image:20080924-emergency_vector_pRLBB_vector_insert_PCR_redo.annotated.jpg|thumb|right|making x6 Parallel MCS Replacement fragmenets via PCR Redo!!! with clean uncontaminated primers [[Media:20080924-emergency_vector_pRLBB_vector_insert_PCR_redo.jpg|download unannotated]]]]
:<strong>Norman</strong>
:<strong>Norman</strong>

Latest revision as of 09:31, 26 September 2008

Projects Events Resources
Sponsors Experiments Milestones Protocols
Notebook (t) Meetings (t)

Things we did today

Wetlab work

Verified plasmid preps

Grace
  • PCR of plasmid preps from yesterday to verify inserts
  • Ran PCR products on gel
  • nrg #1 and nrsg #2 returned blanks. Bad plasmid prep?
  • BB-pRL1383a and colony PCR of BB-pRL1383a E. coli both blank. Yikes!
  • Quantified BB-pRL1383a and pSB1A3 plasmid preps
  • Blanks. Really low conc.? Will nanodrop to check.

Construction of secretion device (cont.)

Grace
  • Overnight RE digest:
  • nir, nrsg, and nrg PCR products with EcoRI and SpeI
  • rgt PCR products with XbaI and PstI
  • J33207 PCR product and pRL1383a plasmid with HindIII and BamHI

Prep for sequencing

Grace
  • nrg #4, nrg #7, nrsg #8, rgt #1, rgt #2
  • plac+rbs
  • PCR machine froze while ExoSAPing. Unable to retrieve samples from machine. Jamie says she will figure it out.

Making Emergency BB Vector Insert (help contaminant troubleshoot)

making x6 Parallel MCS Replacement fragmenets via PCR Redo!!! with clean uncontaminated primers download unannotated
Norman
  • Cross posted in 2008-09-23 for comparison with contaminated PCR gel run.

Discussion

Quote of the Day

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson


Sponsor_UHM.gifSponsor_OVCRGE.gifSponsor_CTAHR.gif