IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-8-24: Difference between revisions

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===Plans===
===Plans===
* Since the Fb used in the assay last night was the only barrel not folded yesterday, and because all the barrels folded at the same time as Fb (ie. [[IGEM:Harvard/2006/DNA_nanostructures/Notebook/2006-8-22#Gel|8.22.06]]) [[IGEM:Harvard/2006/DNA_nanostructures/Notebook/2006-8-22#Gel|looked somewhat misfolded]], it's possible that something went wrong during that folding.  
* Since the Fb used in the assay last night was the only barrel not folded yesterday, and because all the barrels folded at the same time as Fb (ie. [[IGEM:Harvard/2006/DNA_nanostructures/Notebook/2006-8-22#Gel|8.22.06]]) [[IGEM:Harvard/2006/DNA_nanostructures/Notebook/2006-8-22#Gel|looked somewhat misfolded]], it's possible that something went wrong during that folding.  
** Thus, before remaking a large number of pre-working and working stocks and possibly thus wasting materials, I'll first refold using remade Fb (from [[IGEM:Harvard/2006/DNA_nanostructures/Notebook/2006-8-23#Folded:|yesterday]) and retry the assay using just it and c5.0.A.
** Thus, before remaking a large number of pre-working and working stocks and possibly thus wasting materials, I'll first refold using remade Fb (from [[IGEM:Harvard/2006/DNA_nanostructures/Notebook/2006-8-23#Folded:|yesterday]]) and retry the assay using just it and c5.0.A.


===Folded===
===Folded===

Revision as of 13:26, 24 August 2006

PEG fractionation results

Streptavidin Depletion Assay: c5.0.Fb (inside biotinylated barrel)

Plans

  • Since the Fb used in the assay last night was the only barrel not folded yesterday, and because all the barrels folded at the same time as Fb (ie. 8.22.06) looked somewhat misfolded, it's possible that something went wrong during that folding.
    • Thus, before remaking a large number of pre-working and working stocks and possibly thus wasting materials, I'll first refold using remade Fb (from yesterday) and retry the assay using just it and c5.0.A.

Folded

  • 2 rxns of Fb at 30mM MgCl2, using Fb made [[IGEM:Harvard/2006/DNA_nanostructures/Notebook/2006-8-23#Folded:|yesterday].
  • (Additionally, folded for Katie 5 rxns each of:
    • c5.0.A (lidless)
    • c5.0.C (core)
    • c5.0.D (core))

Strep Treatment Protocol

  • Same as that done yesterday, only with just c5.0.A and c5.0.Fb folded today.

Gel

Lane Component Test Condition Amount
1 1kb+ ladder - 10uL
2 p7308 (~42nM) - 9uL
3 c5.0.A barrel (lidless) Untreated 10uL
4 inside biotinylated barrel Untreated 10uL
5 barrel (lidless) Beaded ~38uL
6 inside biotinylated barrel Beaded ~38uL
7 barrel (lidless) Free-strep ~38uL
8 inside biotinylated barrel Free-strep ~38uL
9 barrel (lidless) Free, bead ~38uL
10 inside biotinylated barrel Free, bead ~38uL

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