IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-8-14: Difference between revisions
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==Microcon w/ detergent== | ==Microcon w/ detergent== | ||
* add 20 {{ul}} given nanostructure to center of YM-50 Micrcon tube | |||
* add 480 {{ul}} given folding buffer, microcentrifuge for 6 min. at 14k rcf, and repeat dilution and spinning 4 more times | |||
{| {{table}} | |||
| align="center" style="background:#f0f0f0;"|'''trial''' | |||
| align="center" style="background:#f0f0f0;"|'''nanostructures''' | |||
| align="center" style="background:#f0f0f0;"|'''wash buffer''' | |||
|- | |||
| 6hb-0||20 {{ul}} 6hb||1x folding buffer (10 mM {{mgcl2}}) | |||
|- | |||
| 6hb-0.1||20 {{ul}} 6hb||1x folding buffer (10 mM {{mgcl2}}) w/ 0.1% SDS | |||
|- | |||
| 4-0||20 {{ul}} 4.0.I||1x folding buffer (10 mM {{mgcl2}}) | |||
|- | |||
| 4-0.01||20 {{ul}} 4.0.I||1x folding buffer (10 mM {{mgcl2}}) w/ 0.01% SDS | |||
|- | |||
| 4-0.1||20 {{ul}} 4.0.I||1x folding buffer (10 mM {{mgcl2}}) w/ 0.1% SDS | |||
|} | |||
Revision as of 10:45, 14 August 2006
Goals for today
Microcon Purification Tweaking
- repeat Friday's mega PEG ppt on 5.0 (?)
- Micron experiments with 0.1% and 0.01% SDS in buffer
- ...and use 1x folding buffer and not water for washes
- also: perform control expt with 10 bp+ ladder, since according to Millipore documentation, the filter should retain ds DNAs longer than 100 bp
Streptavidin-Bead "Protection" Assay on Inside- and Outside-Biotinylated c5.0
- NB: no good purification of nanostructure from oligo has been achieved, but gel separation after elution should differentiate formerly bead-bound oligos from formerly bead-bound nanostructures
Redux of [Mg++], [oligos]
- Based on the inconclusive gels from last Monday, the titration will be redone with c5.0 at 8% PEG, 0.5M NaCl final.
Microcon w/ detergent
- add 20 μL given nanostructure to center of YM-50 Micrcon tube
- add 480 μL given folding buffer, microcentrifuge for 6 min. at 14k rcf, and repeat dilution and spinning 4 more times
| trial | nanostructures | wash buffer |
| 6hb-0 | 20 μL 6hb | 1x folding buffer (10 mM MgCl2) |
| 6hb-0.1 | 20 μL 6hb | 1x folding buffer (10 mM MgCl2) w/ 0.1% SDS |
| 4-0 | 20 μL 4.0.I | 1x folding buffer (10 mM MgCl2) |
| 4-0.01 | 20 μL 4.0.I | 1x folding buffer (10 mM MgCl2) w/ 0.01% SDS |
| 4-0.1 | 20 μL 4.0.I | 1x folding buffer (10 mM MgCl2) w/ 0.1% SDS |
