T4 DNA Polymerase

Reagents Supplied

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M0203S     -20       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
M0203L     -20       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
• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)

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. Can T4 DNA Polymerase be used in other NEBuffers and rCutSmart Buffer?
2. Can T4 DNA Polymerase be used to blunt DNA?
3. Can T4 DNA Polymerase be used to fill in 3' overhangs?
4. Can T4 DNA Polymerase be used to remove 5' overhangs?
5. Can T4 DNA Polymerase be heat inactivated?
6. Are the nucleotides needed to remove a 3' overhang using T4 DNA Polymerase?
7. What are the main causes of blunting reaction failure using T4 DNA Polymerase?
8. Can T4 DNA Polymerase be used in labeling reactions and partial fill in reactions?
9. Is T4 DNA Polymerase active at room temperature?
10. Is T4 DNA Polymerase the enzyme of choice for removing 3' overhangs and filling in 5' overhangs (3' recessed ends)?
11. Are NEB DNA Polymerases supplied with dNTPs?
12. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

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.

Specifications & Change Notifications

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
• M0203S_L_v1_0401712
• M0203S_v1_10011307
• M0203S_v1_10021832
• M0203L_v1_10019934
• M0203L_v1_10023074
• M0203S_v1_10029314
• M0203L_v1_10034499
• M0203S_v1_10034498
• M0203L_v1_10036401
• M0203L_v1_10043962
• M0203S_v1_10043963
• M0203L_v1_10048109
• M0203S_v1_10049314
• M0203S_v1_10054179
• M0203L_v1_10055853
• M0203S_v1_10055855
• M0203L_v1_10060120
• M0203S_v1_10060122
• M0203L_v1_10063501
• M0203S_v1_10063553
• M0203L_v1_10068639
• M0203S_v1_10070264
• M0203L_v1_10074951
• M0203S_v1_10075027
• M0203S_v1_10081947
• M0203S_v1_10085012
• M0203L_v1_10085011
• M0203S_v1_10091124
• M0203S_v1_10091904
• M0203L_v1_10091907
• M0203L_v1_10099938
• M0203S_v1_10099937
• M0203L_v1_10107643
• M0203S_v1_10107644
• M0203S_v1_10112939
• M0203L_v1_10113492
• M0203L_v1_10127968
• M0203S_v1_10127969
• M0203S_v1_10143141
• M0203L_v1_10145960
• M0203L_v1_10151704
• M0203S_v1_10151890
• M0203L_v1_10164996
• M0203S_v1_10168847
• M0203L_v1_10179257

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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Licenses

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