Jan 15, 2025
Version 4
Prepare Tris Buffer for Meat identification Protocol V.4
This protocol is a draft, published without a DOI.
- 1OBS
Protocol Citation: openbioscience Adrian 2025. Prepare Tris Buffer for Meat identification Protocol. protocols.io https://protocols.io/view/prepare-tris-buffer-for-meat-identification-protoc-ds826hyeVersion created by openbioscience Adrian
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: In development
We are still developing and optimizing this protocol
Created: September 30, 2024
Last Modified: January 15, 2025
Protocol Integer ID: 113658
Abstract
TRIS buffer protocol for meat identification experiment.
Materials
reagents: Tris, HCl (12.1M), dd-di-DNAase free water. Note: the dilution of HCL can also be 1M.
devices: scale, thermometer, tube rack, pipette rack, pen. optional: incubator, magnetic stirrer or glass rod for stirring, pipettes
labware: 50 ml beaker large enough to permit pH meter measurements
consumables: 50 ml (large) falcon tube
Before start
Make sure you adjust the pH to the same temperature for the reaction in which the buffer is used.
Prepare Tris Buffer for Meat identification Protocol. Prepare (50 ml) of 0.04 mol/L Tris HCl (pH 7.75)
Prepare Tris Buffer for Meat identification Protocol. Prepare (50 ml) of 0.04 mol/L Tris HCl (pH 7.75)
Add 25 mL of de-ionized DNase free water to a 50 ml beaker.
Measure and add 0.24 g of TrisBio Basic Inc.Catalog #TB0196
to the beaker.
Calculation for TrisBio Basic Inc.Catalog #TB0196 weight:
MW for TRIS (C4H11NO3 ) 121.14 mol/Liter
0.04 M TRIS TRIS : m=CMV =0.04mol/L*121.14g/mol *50 ml=0.24 g of TRIS
Heat the beaker to 25 °C Celsius and maintain. This is for accurate pH measurement. Can use a water bath or an incubator to maintain the temp to 25 Celsius for example.
Add about 100 µL or less HCl 12.1 Molarity (M) Hydrochloric AcidBioshopCatalog #HCL333.500 then iteratively measure pH and add 1 to 2μl HCL until pH decreases to 7.75 . The calculation below indicates that about 116uL HCl should achieve the desired pH.
!! All this steps are done at the desired temperature set in the previous step. That is required because any pH measurement has to be done at a set temperature as pH is temperature dependent.
Note
The pH indicated in the referenced paper is 7.75 compared to 7 in the main paper.
Note
The initial pH should be high around 10 or 11 as Tris is alkaline and should decrease with the addition of HCl.
Transfer from beaker to a new 50 ml falcon tube labeled Tris Buffer for storage.
Add water to bring the total to 50 mL .
Store the falcon tube in the fridge for max 4 months.
Protocol references
Normally we make half the solution and titrate with HCl or TRIS-HCl
https://www.researchgate.net/post/How-to-make-a-Tris-HCl-buffer#:~:text=Tris%2DHCl%20can%20be%20prepared,is%20157.6%20g%2Fmol).
pH calculations= https://www.youtube.com/watch?v=387t-nh0OMg
https://www.cusabio.com/m-296.html#a21 for calculating Tris HCL pH from Tris and Hydrocloric acid
https://www.reddit.com/r/labrats/comments/193ei6n/trying_to_prepare_trishcl_buffer_ph_seems/?rdt=42146