IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-8-14

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(Microcon w/ detergent)
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==Microcon w/ detergent==
==Microcon w/ detergent==
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* add 20 {{ul}} given nanostructure to center of YM-50 Micrcon tube
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* add 480 {{ul}} given folding buffer, microcentrifuge for 6 min. at 14k rcf, and repeat dilution and spinning 4 more times
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{| {{table}}
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| align="center" style="background:#f0f0f0;"|'''trial'''
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| align="center" style="background:#f0f0f0;"|'''nanostructures'''
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| align="center" style="background:#f0f0f0;"|'''wash buffer'''
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|-
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| 6hb-0||20 {{ul}} 6hb||1x folding buffer (10 mM {{mgcl2}})
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|-
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| 6hb-0.1||20 {{ul}} 6hb||1x folding buffer (10 mM {{mgcl2}}) w/ 0.1% SDS
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|-
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| 4-0||20 {{ul}} 4.0.I||1x folding buffer (10 mM {{mgcl2}})
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|-
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| 4-0.01||20 {{ul}} 4.0.I||1x folding buffer (10 mM {{mgcl2}}) w/ 0.01% SDS
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|-
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| 4-0.1||20 {{ul}} 4.0.I||1x folding buffer (10 mM {{mgcl2}}) w/ 0.1% SDS
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|}

Revision as of 13:45, 14 August 2006

Contents

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]


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-020 μL 6hb1x folding buffer (10 mM MgCl2)
6hb-0.120 μL 6hb1x folding buffer (10 mM MgCl2) w/ 0.1% SDS
4-020 μL 4.0.I1x folding buffer (10 mM MgCl2)
4-0.0120 μL 4.0.I1x folding buffer (10 mM MgCl2) w/ 0.01% SDS
4-0.120 μL 4.0.I1x folding buffer (10 mM MgCl2) w/ 0.1% SDS
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