T4 DNA Polymerase

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
NEB # Stored at (°C) M0203S -20 Component Name Component # Stored at (°C) Amount Concentration T4 DNA Polymerase M0203SVIAL -20 1 x 0.05 ml 3,000 units/ml NEBuffer™ r2.1 B6002SVIAL -20 1 x 1.25 ml 10 X
NEB # Stored at (°C) M0203L -20 Component Name Component # Stored at (°C) Amount Concentration T4 DNA Polymerase M0203LVIAL -20 1 x 0.25 ml 3,000 units/ml NEBuffer™ r2.1 B6002SVIAL -20 1 x 1.25 ml 10 X

Application Features

• 3´ overhang removal to form blunt ends (1,2).
• 5´ overhang fill-in to form blunt ends (1,2).
• Single strand deletion subcloning (3).
• Second strand synthesis in site-directed mutagenesis (4).
• Probe labeling using replacement synthesis (1,2).

Companion Products

• Deoxynucleotide (dNTP) Solution Set
• Deoxynucleotide (dNTP) Solution Mix

Materials Sold Separately

• NEBuffer™ r2.1

1. For fill-in reactions only: T4 DNA Polymerase can be used in NEBuffers 1.1, 2.1, 3.1 and CutSmart^® Buffer as well as NEBuffers 1-4 and T4 DNA Ligase Reaction Buffer.
2. For blunting reactions requiring removal of overhangs: T4 DNA Polymerase can be used in NEBuffers 1.1, 2.1, and CutSmart Buffer as well as NEBuffers 1, 2, and 4 and T4 DNA Ligase Reaction Buffer. NEBuffers 3.1 and 3 are not recommended when overhang removal is required.
3. Optimal activity is observed in NEBuffer 2.1.
4. Supplement with dNTPs.*
5. BSA supplementation is recommended when using a buffer that does not already contain BSA.
6. Incubate at temperature suggested for specific protocol.
   
   * Refer to specific protocol to determine recommended dNTP concentrations.
   Refer
7. Elevated temperatures, excessive amounts of enzyme, failure to supplement with dNTPs or long reaction times may result in recessed ends due to the 3´→ 5´ exonuclease activity of the enzyme.

1. Tabor, S. and Struhl, K. (1989). DNA-Dependent DNA Polymerases. In F. M. Ausebel, R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith and K. Struhl(Ed.), Current Protocols in Molecular Biology. 3.5.10-3.5.12. New York: John Wiley & Sons, Inc.
2. Sambrook, J. et al. (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.), 5.44-5.47. Cold Spring Harbor: Cold Spring Harbor Laboratory Press.
3. Dale, R. et al. (1985). Plasmid. 13, 31-40.
4. Kunkel, T.A. et al. (1987). Methods Enzymol.. 154, 367-382.
5. Panet, A. et al. (1973). Biochemistry. 12, 5045-5050.

1. Protocol for blunting ends by 3’ overhang removal and 3’ recessed (5’ overhang) end fill-in using T4 DNA Polymerase (M0203)

• Activity of DNA Modifying Enzymes in rCutSmart™ Buffer

• Blunting Selection Chart
• Common Applications for Exonucleases and Endonucleases
• DNA Polymerase Selection Chart

• NEBcloner^®

1. Which DNA polymerase can I use to blunt DNA?
2. Can I use a DNA polymerase to fill in 3' overhangs or remove 5' overhangs?
3. How can I heat inactivate this DNA polymerase?
4. Can this DNA polymerase be used in labeling and partial fill reactions?
5. What are the main causes of blunting reaction failure?
6. Can this DNA polymerase be used in other NEBuffers for filling in or blunting overhangs?

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]

• M0203S_L_v1

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]

• M0203S_L_v1_0401512
• M0203S_L_v1_0401606
• M0203S_L_v1_0401612
• M0203S_L_v1_0401706
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• M0203S_v1_10234129
• M0203S_v1_10240886
• M0203L_v1_10243044
• M0203S_v1_10255804
• M0203L_v1_10255851
• M0203S_v1_10260747
• M0203L_v1_10269205
• M0203S_v1_10270903
• M0203S_v1_10281956
• M0203L_v1_10287139
• M0203S_v1_10290032
• M0203S_v1_10300230
• M0203L_v1_10300309
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• M0203L_v1_10327137
• M0203S_v1_10338955
• M0203L_v1_10342316

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

• T4 DNA Polymerase
• NEBuffer™ r2.1

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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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