Feb 20, 2022

Public workspaceRadioactive Labeling with T4 PNK 3' phosphatase minus (M0236) V.2

DOI
dx.doi.org/10.17504/protocols.io.bd23i8gn
  • 1New England Biolabs
  • New England Biolabs (NEB)
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DOI: dx.doi.org/10.17504/protocols.io.bd23i8gn
External link: https://www.neb.com/protocols/2012/08/24/radioactive-labeling-with-t4-pnk-or-t4-pnk-3-phosphatase-minus
Protocol Citation: New England Biolabs 2022. Radioactive Labeling with T4 PNK 3' phosphatase minus (M0236). protocols.io https://dx.doi.org/10.17504/protocols.io.bd23i8gnVersion created by New England Biolabs
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: March 21, 2020
Last Modified: February 20, 2022
Protocol Integer ID: 34619
Keywords: T4PNK Reaction Buffer, radiolabeling, Radioactive , Labeling, T4, PNK, T4 Polynucleotide Kinase
Abstract
Radioactive Labeling with T4 Polynucleotide Kinase 3' phosphatase minus.
Materials
MATERIALS
ReagentT4 Polynucleotide Kinase (3' phosphatase minus) - 200 unitsNew England BiolabsCatalog #M0236S
Safety warnings
Please refer to the Safety Data Sheets (SDS) for health and environmental hazard.
Set-up the following reaction:
AB
ComponentAmount
DNAUp to 50 pmol of 5´ termini
T4 PNK Reaction Buffer (10X)5 µl
ATP (10 mM)50 pmol of [γ-32P] ATP
T4 PNK2 µl (20 units)
Nuclease-free Waterup to 50 µl

Note
Note: [33P] ATP may be substituted for [32P] ATP.

Pipetting
Incubate at Temperature37 °C for Duration00:30:00 .
Incubation
Heat inactivate by incubating at Temperature65 °C for Duration00:20:00 .
Incubation