May 11, 2017

Public workspaceResuspension Buffer P1 V.2

DOI
https://dx.doi.org/10.17504/protocols.io.hxgb7jw
  • Qiagen 1
  • 1Matthew Sullivan Lab, University of Arizona, The Ohio State University
  • VERVE Net
  • Sullivan Lab
Bonnie Poulos
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DOI: https://dx.doi.org/10.17504/protocols.io.hxgb7jw
External link: http://people.mbi.ucla.edu/sumchan/qiagenbuffer.html
Protocol Citation: Qiagen 2017. Resuspension Buffer P1. protocols.io https://dx.doi.org/10.17504/protocols.io.hxgb7jw
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
Created: May 11, 2017
Last Modified: April 13, 2018
Protocol Integer ID: 5832
Keywords: resuspension buffer p1, containing rnase, ph, cl, edta
Abstract
This is a 0.05M Tris-Cl (pH 8.0), 0.01M EDTA resuspension buffer containing RNase A
Guidelines
Storage condition - 4°C after adding RNase A.
Note: Buffer compositions are given per liter of solution. Buffer calculations are based on Tris Base adjusted to pH 8.0 with HCl (Tris-Cl).  Not recommended to be made with Tris-HCl due to acidity of solution (EDTA-Na2-2H2O needs a pH above 8.0 to dissolve), but if Tris-HCl is used, amount should be adjusted for correct molarity (0.05M) and NaOH should be used to bring pH up to 8.0.  This buffer may be autoclaved prior to addition of RNase A or it may be 0.2 μm filter-sterilized.
Materials
STEP MATERIALS
ReagentTris BaseFisher ScientificCatalog #BP152-1
ReagentEDTA, disodium salt, dihydrateFisher ScientificCatalog #S312-500
ReagentTris BaseFisher ScientificCatalog #BP152-1
ReagentEDTA, disodium salt, dihydrateFisher ScientificCatalog #S312-500
Protocol materials
ReagentTris BaseFisher ScientificCatalog #BP152-1
ReagentEDTA, disodium salt, dihydrateFisher ScientificCatalog #S312-500
Troubleshooting
Dissolve 6.06g Tris Base in 800 mL MilliQ water
Amount6 µL
Note
It can be made either way depending on what you have available in the lab. Some find it easier to use stock solutions, but it is not necessary as long as final pH is adjusted.
ReagentTris BaseFisher ScientificCatalog #BP152-1
Add 3.72g EDTA disodium salt, dihydrate to the 800 mL Tris base and stir to dissolve
Amount4 µL
ReagentEDTA, disodium salt, dihydrateFisher ScientificCatalog #S312-500
Adjust the pH to 8.0 with HCl
Adjust the volume to 1 liter with MilliQ water
Add 100mg RNase A per liter of buffer P1
Amount100 µL