Nov 15, 2020

Public workspaceNEBNext Ultra End Prep Mixture E7442 V.4

DOI
dx.doi.org/10.17504/protocols.io.bpiumkew
  • 1New England Biolabs
  • New England Biolabs (NEB)
    Tech. support phone: +1(800)632-7799 email: info@neb.com
Isabel Gautreau
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DOI: dx.doi.org/10.17504/protocols.io.bpiumkew
External link: https://www.neb.com/protocols/2012/11/15/nebnext-end-prep-e7370
Protocol Citation: Isabel Gautreau 2020. NEBNext Ultra End Prep Mixture E7442. protocols.io https://dx.doi.org/10.17504/protocols.io.bpiumkewVersion created by Isabel Gautreau
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: November 09, 2020
Last Modified: November 15, 2020
Protocol Integer ID: 44340
Materials
MATERIALS
ReagentNEBNext End Prep Enzyme MixNew England BiolabsCatalog #E7646
ReagentNEBNext End Repair Reaction BufferNew England BiolabsCatalog #E7372
STEP MATERIALS
ReagentNEBNext End Prep Enzyme MixNew England BiolabsCatalog #E7646
ReagentNEBNext End Repair Reaction BufferNew England BiolabsCatalog #E7372
ReagentNEBNext End Prep Enzyme MixNew England BiolabsCatalog #E7646
ReagentNEBNext End Repair Reaction BufferNew England BiolabsCatalog #E7372
Protocol materials
ReagentNEBNext End Repair Reaction BufferNew England BiolabsCatalog #E7372
ReagentNEBNext End Prep Enzyme MixNew England BiolabsCatalog #E7646
ReagentNEBNext End Repair Reaction BufferNew England BiolabsCatalog #E7372
ReagentNEBNext End Prep Enzyme MixNew England BiolabsCatalog #E7646
ReagentNEBNext End Repair Reaction BufferNew England BiolabsCatalog #E7372
ReagentNEBNext End Prep Enzyme MixNew England BiolabsCatalog #E7646
ReagentNEBNext End Prep Enzyme MixNew England BiolabsCatalog #E7646
ReagentNEBNext End Repair Reaction BufferNew England BiolabsCatalog #E7372
NEBNext End Prep
NEBNext End Prep
Starting Material: 5 ng-1 μg fragmented DNA
Mix the following comonents in a sterile, nuclease-free tube:
ComponentVolume
End Prep Enzyme Mix3 µl
End Repair Reaction Buffer (10X)6.5 µl
Fragmented DNA55.5 µl
Total Volume65 µl
ReagentNEBNext End Prep Enzyme MixNew England BiolabsCatalog #E7646
ReagentNEBNext End Repair Reaction BufferNew England BiolabsCatalog #E7372
Mix by pipetting, followed by a quick spin to collect all liquid from the sides of the tube.
Place in a thermocycler, with the heated lid on, and run the following program:
30 minutes @ 20°C
30 minutes @65°C
Hold at 4°C
Proceed directly to NEBNext Ultra Ligation Module (NEB #E7445).