Oct 01, 2024
Version 1
Prepare Tris Buffer for Meat identification Protocol V.1
- 1OBS
Protocol Citation: openbioscience Adrian 2024. Prepare Tris Buffer for Meat identification Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.kqdg32z1zv25/v1
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
Obsolete
Created: October 01, 2024
Last Modified: October 01, 2024
Protocol Integer ID: 108692
Abstract
Buffer protocol for meat identification experiment
Materials
reagents: Tris, HCl (12.1M), dd-di-DNAase free water
devices: scale, thermometer optional: incubator, magnetic stirrer or glass rod for stirring
consumables: 50 ml tube
Protocol materials
TrisBio Basic Inc.Catalog #TB0196
Hydrochloric AcidBioshopCatalog #HCL333.500
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 deionized DNAse free, dd water to a 50 ml falcon tube. Bring to to final experiment desired Celsius temperature (22 Celsius for example) and maintain. Can use an incubator for example.
Measure and add 0.24 g of Tris
Calculation for TrisBio Basic Inc.Catalog #TB0196 weight:
MW for TRIS Hydrochloride (C4H11NO3 ·HCl) 121.14 mol/Liter, (Hydrochloric HCl Acid, MW: 36.46 g/mol)
0.04 M TRIS TRIS Hydrochloride: m=CMV =0.04mol/L*121.14g/mol *50 ml=0.24 g of TRIS
Add 100 μl HCl 12.1M
Hydrochloric AcidBioshopCatalog #HCL333.500
iteratively measure pH and add 1 to 2μl HCL(12M) until pH reaches 7.75. Calculator indicates 116.
Add water to 50 ml
Store in the fridge for max 4 months
Protocol references
Normally we make half the solution and titrate with 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