Jun 14, 2022
Production: 10x TBE Buffer Powder Sachets
This protocol is a draft, published without a DOI.
- 1Beneficial Bio;
- 2Beneficial Bio, Mboalab
Protocol Citation: Jenny Molloy, Nadine Mowoh, Stephane Fadanka 2022. Production: 10x TBE Buffer Powder Sachets. protocols.io https://protocols.io/view/production-10x-tbe-buffer-powder-sachets-b94ur8ww
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: May 27, 2022
Last Modified: June 14, 2022
Protocol Integer ID: 63348
Abstract
Desired concentrations of Components in the 10x buffer
Boric acid-0.89M
Tris-0.89M
Na2EDTA-0.02M
Concentration of 1x TBE buffer
Tris-Borate-0.089M
Na2EDTA-0.002M
Materials
BoraxContributed by users
Tris baseContributed by users
EDTA DiSodium salt anhydrateSigma Aldrich
Weighing balance and weighing boat
Spatula
Ziplock Bags
Protocol materials
Borax
Materials, Step 4
Tris base
Materials, Step 6
EDTA DiSodium salt anhydrateMerck MilliporeSigma (Sigma-Aldrich)
Materials, Step 5
Safety warnings
Work in well ventilated area and weigh face masks and googles to avoid inhaling the powders or powder from entering the eye.
Before start
Ensure all powders are dry and free from moisture to enable the resulting buffer to stay longer.
Weigh about 35 g of Boric acid into a weighing boat and place the weighing boat in a vacuum tupperware half-filled with silica gel beads.
5m
Close the tupperware and use the vacuum pump to create a vacuum until you are not able to hand pump any more air out or the pump shuts off.
Note
Check instructions for your vacuum pump system to understand when a vacuum has been formed and when to stop pumping.
2m
Allow it to stay overnight at room temperature
Note
This step ensures no moisture is carried over into the final powder mixture. The powders readily absorb moisture from the atmosphere and this can reduce their shelf life.
12h
Weigh out 27.515 g of Borax from the pre-dried powder into a 500 mL beaker.
5m
Weigh 3.72 g of EDTA DiSodium salt anhydrateSigma Aldrich and add into the beaker
2m
Weigh 53.905 g ofTris base and add to the content of the beaker.
2m
Use a spatula to stir so as to have an even and a uniform mixture of all three powders.
Note
If scaling up the protocol, stirring may be insufficient and you may want to vigorously agitate in a close container to ensure that the powders are evenly mixed.
2m
Transfer the powder mix into a zip lock antistatic bag, seal carefully and label appropriately as powder to make up 0.5 L of 10x TBE.
2m
The powder is stable for up to 6 months.
Note
For long term storage, keep the pouches in a secondary container like a larger tupperware box with dessicant to help absorb any extra moisture from the atmosphere.