T4 Polynucleotide Kinase

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M0201S     -20       T4 Polynucleotide Kinase M0201SVIAL -20 1 x 0.05 ml 10,000 units/ml   T4 Polynucleotide Kinase Reaction Buffer B0201SVIAL -20 1 x 1 ml 10 X
M0201L     -20       T4 Polynucleotide Kinase M0201LVIAL -20 1 x 0.25 ml 10,000 units/ml   T4 Polynucleotide Kinase Reaction Buffer B0201SVIAL -20 1 x 1 ml 10 X

Application Features

• End-labeling DNA or RNA for probes and DNA sequencing (2)
• Addition of 5´-phosphates to oligonucleotides to allow subsequent ligation
• Removal of 3´-phosphoryl groups (3)

Materials Sold Separately

• T4 Polynucleotide Kinase Reaction Buffer

1. Gaps can be phosphorylated with elevated levels of ATP. Nicks are not phosphorylated efficiently. CTP, GTP, TTP, UTP, dATP or dTTP can be substituted for ATP as a phosphate donor (1).
2. Dephosphorylation prior to end-labeling can be avoided by utilizing the exchange reaction, although this results in lower specific activity labeling.
3. Sufficient incorporation levels can be attained using the supplied buffer supplemented with 100 µM ADP in the final reaction. However, higher levels of incorporation with the exchange reaction are achieved when using the buffer described in (2).

1. Richardson, CC. (1981). The Enzyme. 14, 299-314.
2. Sambrook, J. et al. (1989). Molecular Cloning: A Laboratory Manual.. 10.59-10.67,11.31-11.33.
3. Cameron, V. and Uhlenbeck, O.C. (1977). Biochemistry. 16, 5120-5126.
4. Berkner, K.L. and Folk, W.R. (1977). J. Biol. Chem.,. 252, 3176-3184.
5. Soltis, D.A. and Uhlenbeck, 0.C. (1982). J. Biol. Chem.. 257, 11332-11339.
6. Wang, L.K. and Human, S. (2001). J. Biol. Chem.. 276, 26868-26874.
7. Wang, L.K. and Shuman, S. (2002). Nucl. Acids Res.. 30, 1073-1080.
8. Vance, J.R. and Wilson, T.E. (2001). Mol. Cell. Biol. 21, 7191-7198.
9. Interthal, H. et al. (2005). J. Biol. Chem.,. 280, 36518-36528.

1. Radioactive Labeling with T4 PNK or T4 PNK (3´ phosphatase minus)
2. Non-radioactive Phosphorylation with T4 PNK or T4 PNK (3´ phosphatase minus)
3. End-labeling Protocol

• Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

• NEB Diluent and Buffer Table

• NEBcloner^®

1. What factors can cause incomplete phosphorylation when using T4 Polynucleotide Kinase?
2. Can I use T4 Polynucleotide Kinase and T4 DNA Ligase in the same reaction buffer?
3. How can the rate of phosphorylation be improved when using T4 Polynucleotide Kinase?
4. Do I need to dephosphorylate prior to labeling?
5. How much substrate can be phosphorylated in a standard reaction?
6. How many units of T4 Polynucleotide Kinase should be used for a typical reaction?
7. How do I inactivate the enzyme?
8. How to calculate the molarity of ends?
9. What labels can be used?
10. Will T4 Polynucleotide Kinase work in rCutSmart Buffer?

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

• M0201S_L_v2

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

• M0201S_L_v1_0951607
• M0201S_L_v1_0951703
• M0201S_L_v1_0951711
• M0201S_v1_10008062
• M0201L_v1_10008060
• M0201S_v1_10014711
• M0201S_v1_10015098
• M0201L_v1_10016592
• M0201S_v1_10018038
• M0201S_v1_10020295
• M0201S_v1_10025785
• M0201L_v1_10025784
• M0201S_v1_10028772
• M0201S_v1_10031155
• M0201L_v1_10028773
• M0201S_v1_10031197
• M0201L_v1_10032258
• M0201S_v1_10032256
• M0201S_v1_10033445
• M0201S_v1_10039230
• M0201L_v1_10039231
• M0201S_v1_10042537
• M0201S_v1_10045042
• M0201L_v1_10045208
• M0201S_v1_10045210
• M0201S_v1_10048640
• M0201S_v1_10051581
• M0201L_v1_10051580
• M0201S_v1_10053804
• M0201L_v1_10057949
• M0201S_v1_10057256
• M0201L_v1_10061779
• M0201S_v1_10061778
• M0201S_v1_10064351
• M0201S_v1_10066281
• M0201L_v1_10065600
• M0201S_v1_10068950
• M0201L_v1_10070008
• M0201S_v1_10070007
• M0201S_v1_10079367
• M0201L_v1_10081113
• M0201S_v1_10082467
• M0201L_v1_10082468
• M0201S_v1_10085461
• M0201S_v1_10088982
• M0201S_v1_10092452
• M0201S_v1_10093429
• M0201L_v1_10092451
• M0201S_v1_10094759
• M0201L_v1_10097689
• M0201S_v1_10098674
• M0201S_v1_10102420
• M0201L_v1_10102419
• M0201S_v1_10108414
• M0201L_v1_10110944
• M0201S_v1_10111493
• M0201L_v1_10111494
• M0201S_v1_10115995
• M0201L_v1_10115996
• M0201S_v1_10120089
• M0201L_v1_10126413
• M0201S_v1_10126412
• M0201L_v1_10129474
• M0201S_v1_10129473
• M0201S_v1_10138992
• M0201L_v1_10138993
• M0201S_v1_10145028
• M0201L_v1_10148978
• M0201S_v1_10148980
• M0201S_v1_10156398
• M0201L_v1_10156399
• M0201S_v1_10160735
• M0201L_v1_10165547
• M0201S_v1_10168638
• M0201S_v1_10173718
• M0201L_v1_10173924
• M0201S_v1_10176582
• M0201L_v1_10180293
• M0201S_v1_10185384
• M0201L_v1_10185385
• M0201S_v1_10192172
• M0201L_v1_10195923
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• M0201S_v1_10211629
• M0201L_v1_10218654
• M0201S_v1_10218658
• M0201S_v1_10221771
• M0201L_v1_10221770
• M0201L_v1_10234136
• M0201S_v1_10236793
• M0201S_v1_10238784
• M0201L_v2_10243109
• M0201S_v2_10243110
• M0201S_v2_10252190
• M0201L_v2_10252191
• M0201S_v2_10262424
• M0201L_v2_10263594
• M0201S_v2_10269451
• M0201L_v2_10274397
• M0201S_v2_10280701
• M0201L_v2_10281994
• M0201S_v2_10279056
• M0201L_v2_10281950
• M0201L_v2_10291317
• M0201S_v2_10290033
• M0201S_v2_10300219
• M0201L_v2_10300669
• M0201S_v2_10303225
• M0201L_v2_10310327
• M0201S_v2_10314220
• M0201L_v2_10321113
• M0201S_v2_10324842
• M0201L_v2_10330659
• M0201S_v2_10335956

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

• T4 Polynucleotide Kinase
• T4 Polynucleotide Kinase Reaction Buffer

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This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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