Sep 09, 2020
FloodLAMP Binding Solution Prep v1.1
- Randy True1
- 1FloodLAMP Biotechnologies PBC
- FloodLAMP
- XPRIZE Rapid Covid Testing
Protocol Citation: Randy True 2020. FloodLAMP Binding Solution Prep v1.1. protocols.io https://dx.doi.org/10.17504/protocols.io.bk5sky6e
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: Working
We use this protocol and it's working
Created: September 08, 2020
Last Modified: September 09, 2020
Protocol Integer ID: 41874
Abstract
The Binding Solution includes NaI, HCl, Triton-X100, and water. We have had lots of trouble with it, largely due to a mistake that has been corrected. In our hands currently, the Binding Solution is a pale yellow—a possibility mentioned in the Rabe Cepko paper. It does seem to become cloudy over time (days to weeks). Our current plan is to prepare weighed-out NaI and the other components premixed (stored at 4C) for easier prep, and then make fresh Binding Solution up at least weekly.
We have moved to storing the Binding Solution in 5mL amber screw cap tubes for development runs (2.25mL for a single strip of 8 samples), with these in an opaque container. This makes it convenient to add glass milk and use it as a master mix (as suggested by Rabe Cepko). Care must be taken to keep the silica resuspended prior to pipetting for the addition to the sample, hence the mixing by pipetting on every draw.
Guidelines
Individuals are responsible for the chemical safety training to safely complete this protocol. This procedure should be completed in a chem fume hood with appropriate PPE.
Materials
MATERIALS
Hydrochloric acidSigma AldrichCatalog #H1758
UltraPure Distilled WaterThermo Fisher ScientificCatalog #10977015
Triton X-100Sigma AldrichCatalog #X100-100ML
Sodium Iodide (NaI)SigmaCatalog #793558
- 50mL Falcon tubes
- 5mL amber Eppendorf DNA tube with screw cap ($85 for 200)
150mL Nalgene vacuum Filter with .22um PES filter (12 for $92)
Safety warnings
Hydrochloric acid is very caustic and should be handled with caution. Be extra careful cleaning spills as to not create toxic chlorine gas. Sodium Iodide is hazardous if ingested.
Weight out 45g of NaI into 50mL Falcon Tube
Add 1st traunch of water to 45ml
Vortex, shake and then vortex again until fully mixed
Add 2nd traunch of water to 46mL
Vortex, shake and then vortex again until fully mixed
Vacuum filter (or wait and vacuum filter many tubes at the end)
Add .5mL of 1N HCl
Vortex for 10s, be careful that the tube is completely closed
Add 1.0 mL of Triton X-100, note that it is very viscous (use 1000ul tip with end cut off)
Vortex for 10s and shake
Add 2.5mL UltraPure dH2O
Vortex 10s
Aliquot to 5mL amber Eppendorf screw caps, 2.25mL for 8 x .5mL samples, store in dark box