Klenow Fragment (3'→5' exo-)

Application Features

• Random priming labeling
• DNA sequencing by the Sanger dideoxy method (2)
• Second strand cDNA synthesis
• Second strand synthesis in mutagenesis protocols (3).

Companion Products

• Deoxynucleotide (dNTP) Solution Set
• Deoxynucleotide (dNTP) Solution Mix
• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)

Materials Sold Separately

• NEBuffer 2

1. Klenow Fragment (3´→ 5´ exo-) is not suitable for generating blunt ends because it lacks the 3´→ 5´ exonuclease necessary to remove non-templated 3´ additions.
2. Klenow Fragment (3´→ 5´ exo-) is also active in all four NEBuffers when supplemented with dNTPs.
3. When Klenow Fragment (3´→ 5´ exo-) is used to sequence DNA using the dideoxy method of Sanger et al., 1 unit/5 µl reaction volume is recommended.

1. Derbyshire, V. et al. (1988). Science. 240, 199-201.
2. Sanger, F. et al. (1977). Proc. Natl. Acad. Sci. USA. 74, 5463-5467.
3. Gubler, U. (1987). In S.L. Berger & A.R. Rimmel(Ed.), Methods in Enzymology. 152, 330-335. San Diego: Academic Press.
4. Bebenek, K., Joyce, C.M., Fitzgerald, M.P. and Kunkel, T.A. (1990). J. Biol. Chem.. 265, 13878-13887.

1. A-Tailing with Klenow Fragment (3'-->5' exo-)

• Activity of DNA Modifying Enzymes in CutSmart® Buffer

• A-tailing Selection Chart
• Common Applications for Exonucleases and Endonucleases
• DNA Polymerase Selection Chart

• NEBcloner

1. Can Klenow Fragment (3'→5' exo-) be used in other NEBuffers, including CutSmart?
2. Can Klenow Fragment (3'→5' exo-) be used to blunt DNA?
3. Can Klenow Fragment (3'→5' exo-) be used to fill in 3' overhangs?
4. Can Klenow Fragment (3'→5' exo-) be used to remove 5' overhangs?
5. Can Klenow Fragment (3'→5' exo-) be heat inactivated?
6. Can Klenow Fragment (3'→5' exo-) be used in labeling reactions and partial fill in reactions?
7. Are there other methods for making probes?
8. What is the enzyme of choice for chewing back 3' overhangs and filling in 5' overhangs (3' recessed ends)?
9. Are NEB DNA Polymerases supplied with dNTPs?

Product Citation Tool

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]

• M0212M_v1
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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]

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• M0212L_v2_10083165
• M0212M_v2_10085113
• M0212S_v2_10084994

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

• Klenow Fragment (3'→5' exo-)
• NEBuffer™ 2

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