Sep 07, 2020

Public workspaceGlass Milk preparation - forked

 Forked from Glass Milk preparation
This protocol is a draft, published without a DOI.
  • 1University of Dublin, Trinity College
  • MC2
Martin Codyre
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Protocol Citation: Martin Codyre 2020. Glass Milk preparation - forked. protocols.io https://protocols.io/view/glass-milk-preparation-forked-bkzykx7w
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Other
We attempted this protocol, but could not get it to work in our workspace
Created: September 07, 2020
Last Modified: September 07, 2020
Protocol Integer ID: 41752
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Materials
MATERIALS
ReagentTris, 1 M, pH 8.0AmbionCatalog #AM9855G
Reagent Hydrochloric acidSigma – AldrichCatalog #320331-500ML
ReagentUltraPure™ 0.5 M EDTA pH 8.0Thermo Fisher ScientificCatalog #15575020
ReagentSilica 325 Mesh
STEP MATERIALS
Reagent Hydrochloric acidSigma – AldrichCatalog #320331-500ML
ReagentMilliQ water
ReagentMilliQ water
ReagentTris, 1 M, pH 8.0AmbionCatalog #AM9855G
ReagentUltraPure™ 0.5 M EDTA pH 8.0Thermo Fisher ScientificCatalog #15575020
325 mesh silicon dioxide (Spectrum Chemicals - SI108) Silica 325 mesh is a flint glass powder available from ceramic shops https://www.spectrumchemical.com/OA_HTML/chemical-products_Silicon-Dioxide-325-Mesh-Crystalline_SI108.jsp?section=16930

Millipore Sigma 320331 HCl
Protocol materials
ReagentMilliQ water
ReagentTris, 1 M, pH 8.0AmbionCatalog #AM9855G
Reagent Hydrochloric acidMerck MilliporeSigma (Sigma-Aldrich)Catalog #320331-500ML
ReagentSilica 325 Mesh
Reagent Hydrochloric acidMerck MilliporeSigma (Sigma-Aldrich)Catalog #320331-500ML
ReagentMilliQ water
ReagentTris, 1 M, pH 8.0AmbionCatalog #AM9855G
ReagentUltraPure™ 0.5 M EDTA pH 8.0Thermo Fisher ScientificCatalog #15575020
ReagentUltraPure™ 0.5 M EDTA pH 8.0Thermo Fisher ScientificCatalog #15575020
ReagentSilica 325 Mesh
Reagent Hydrochloric acidMerck MilliporeSigma (Sigma-Aldrich)Catalog #320331-500ML
ReagentMilliQ water
ReagentMilliQ water
ReagentTris, 1 M, pH 8.0AmbionCatalog #AM9855G
ReagentUltraPure™ 0.5 M EDTA pH 8.0Thermo Fisher ScientificCatalog #15575020
Safety warnings
dry silica powder should not be inhaled
Glass Milk Preparation
Glass Milk Preparation
7h
7h
To prepare glass milk, 325 mesh silicon dioxide (Spectrum Chemicals - SI108)
ReagentSilica 325 Mesh
was combined with an excess volume of 10% HCl (~3 N HCl)
made from combining 37%



Reagent Hydrochloric acidMerck MilliporeSigma (Sigma-Aldrich)Catalog #320331-500ML
and MilliQ water (Millipore) in a fume hood
ReagentMilliQ water


Equipment
Fume hood
NAME
Fume hood
TYPE
Generic
BRAND
Unknown
SKU


Safety information


Note
(dry silica powder should not be inhaled).


45m
Sub step in forked version for testing.
30s
After acid washing for Duration04:00:00 Possibly 4 to 8 hours at room temperature TemperatureRoom temperature , silica was pelleted by spinning two minutes at Centrifigation5000 rpm, 00:02:00 5,000 xg and the supernatant was poured off.
6h
The pellet was resuspended in four pellet volumes of
ReagentMilliQ water
and then pelleted again.
This wash step was repeated for a total of six washes.

1h

The pellet was then washed with four pellet volumes of 10 mM Tris HCl, pH = 8 (ThermoFisher Scientific AM9855G)

ReagentTris, 1 M, pH 8.0AmbionCatalog #AM9855G
and 1 mM EDTA (ThermoFisher Scientific 15575020),

ReagentUltraPure™ 0.5 M EDTA pH 8.0Thermo Fisher ScientificCatalog #15575020

and pelleted.
30m
Finally, the pellet was resuspended in 1 pellet volume of 10 mM Tris HCl and 1 mM EDTA and autoclaved. This autoclave step is likely superfluous, however, as acid washes should render the beads free of contaminants. The resulting 50% glass milk slurry can be stored at room temperature.

Before use, care must be taken to vigorously resuspend the particles as they begin to settle quickly.
30m