上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。
T4 DNA Polymerase
We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.
T4 DNA Polymerase catalyzes the synthesis of DNA in the 5´→ 3´ direction and requires the presence of template and primer.
Gap filling (no strand displacement activity)
Removal of 3’ overhangs or fill-in of 5’ overhangs to form blunt ends
Lacks 5’ —> 3’ exonuclease activity
Probe labeling using replacement synthesis
Singe-strand deletion subcloning
精选视频
DNA Blunting Tutorial
观看其他视频
Need a custom/large volume order? Contact Us
Bulk packaging may also be available and requested for large recurring orders. Learn More
产品信息
T4 DNA Polymerase catalyzes the synthesis of DNA in the 5´→ 3´ direction and requires the presence of template and primer. This enzyme has a 3´→ 5´ exonuclease activity which is much more active than that found in DNA Polymerase I (E. coli). Unlike E. coli DNA Polymerase I, T4 DNA Polymerase does not have a 5´→ 3´ exonuclease function.
产品来源
Purified from a strain of E. coli that carries the T4 DNA Polymerase gene.
产品类别:
DNA Manipulation Products
应用:
Blunting,
Polymerases for DNA Manipulation,
PCR
产品组分信息
产品组分信息
本产品提供以下试剂或组分:
NEB #
名称
组分货号
储存温度
数量
浓度
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
特性和用法
单位定义
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C (5).
反应条件
1X NEBuffer™ r2.1
1X NEBuffer™ r2.1 50 mM NaCl 10 mM Tris-HCl 10 mM MgCl2 100 µg/ml Recombinant Albumin (pH 7.9 @ 25°C)
Second strand synthesis in site-directed mutagenesis (4).
Probe labeling using replacement synthesis (1,2).
相关产品
相关产品
Deoxynucleotide (dNTP) Solution Set
Deoxynucleotide (dNTP) Solution Mix
BSA,分子生物学级
Monarch® Plasmid Miniprep Kit
Monarch® DNA Gel Extraction Kit
Monarch® PCR & DNA Cleanup Kit (5 μg)
单独销售的组分
NEBuffer™ r2.1
注意事项
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.
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.
Optimal activity is observed in NEBuffer 2.1.
Supplement with dNTPs.*
BSA supplementation is recommended when using a buffer that does not already contain BSA.
Incubate at temperature suggested for specific protocol.
* Refer to specific protocol to determine recommended dNTP concentrations.
Refer
参考文献
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.
Sambrook, J. et al. (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.), 5.44-5.47. Cold Spring Harbor: Cold Spring Harbor Laboratory Press.
Dale, R. et al. (1985). Plasmid. 13, 31-40.
Kunkel, T.A. et al. (1987). Methods Enzymol.. 154, 367-382.
Panet, A. et al. (1973). Biochemistry. 12, 5045-5050.
操作说明、说明书 & 用法
操作说明
Protocol for blunting ends by 3’ overhang removal and 3’ recessed (5’ overhang) end fill-in using T4 DNA Polymerase (M0203)
使用指南
Activity of DNA Modifying Enzymes in rCutSmart™ Buffer
工具 & 资源
选择指南
Blunting Selection Chart
Common Applications for Exonucleases and Endonucleases
DNA Polymerase Selection Chart
Web 工具
NEBcloner®
FAQs & 问题解决指南
FAQs
Can T4 DNA Polymerase be used in other NEBuffers and rCutSmart Buffer?
Can T4 DNA Polymerase be used to blunt DNA?
Can T4 DNA Polymerase be used to fill in 3′ overhangs?
Can T4 DNA Polymerase be used to remove 5′ overhangs?
Can T4 DNA Polymerase be heat inactivated?
Are the nucleotides needed to remove a 3′ overhang using T4 DNA Polymerase?
What are the main causes of blunting reaction failure using T4 DNA Polymerase?
Can T4 DNA Polymerase be used in labeling reactions and partial fill in reactions?
Is T4 DNA Polymerase active at room temperature?
Is T4 DNA Polymerase the enzyme of choice for removing 3′ overhangs and filling in 5′ overhangs (3′ recessed ends)?
Are NEB DNA Polymerases supplied with dNTPs?
Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
实验技巧
Excess enzyme, temperatures above 12°C, or limited dNTP concentrations can cause excessive 3’ → 5’ exonuclease degradation, which eliminates blunt end formation.
上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。
T4 DNA Polymerase
We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.
T4 DNA Polymerase catalyzes the synthesis of DNA in the 5´→ 3´ direction and requires the presence of template and primer.
Gap filling (no strand displacement activity)
Removal of 3’ overhangs or fill-in of 5’ overhangs to form blunt ends
Lacks 5’ —> 3’ exonuclease activity
Probe labeling using replacement synthesis
Singe-strand deletion subcloning
精选视频
DNA Blunting Tutorial
观看其他视频
Need a custom/large volume order? Contact Us
Bulk packaging may also be available and requested for large recurring orders. Learn More
产品信息
T4 DNA Polymerase catalyzes the synthesis of DNA in the 5´→ 3´ direction and requires the presence of template and primer. This enzyme has a 3´→ 5´ exonuclease activity which is much more active than that found in DNA Polymerase I (E. coli). Unlike E. coli DNA Polymerase I, T4 DNA Polymerase does not have a 5´→ 3´ exonuclease function.
产品来源
Purified from a strain of E. coli that carries the T4 DNA Polymerase gene.
产品类别:
DNA Manipulation Products
应用:
Blunting,
Polymerases for DNA Manipulation,
PCR
产品组分信息
产品组分信息
本产品提供以下试剂或组分:
NEB #
名称
组分货号
储存温度
数量
浓度
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
特性和用法
单位定义
One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C (5).
反应条件
1X NEBuffer™ r2.1
1X NEBuffer™ r2.1 50 mM NaCl 10 mM Tris-HCl 10 mM MgCl2 100 µg/ml Recombinant Albumin (pH 7.9 @ 25°C)
Second strand synthesis in site-directed mutagenesis (4).
Probe labeling using replacement synthesis (1,2).
相关产品
相关产品
Deoxynucleotide (dNTP) Solution Set
Deoxynucleotide (dNTP) Solution Mix
BSA,分子生物学级
Monarch® Plasmid Miniprep Kit
Monarch® DNA Gel Extraction Kit
Monarch® PCR & DNA Cleanup Kit (5 μg)
单独销售的组分
NEBuffer™ r2.1
注意事项
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.
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.
Optimal activity is observed in NEBuffer 2.1.
Supplement with dNTPs.*
BSA supplementation is recommended when using a buffer that does not already contain BSA.
Incubate at temperature suggested for specific protocol.
* Refer to specific protocol to determine recommended dNTP concentrations.
Refer
参考文献
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.
Sambrook, J. et al. (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.), 5.44-5.47. Cold Spring Harbor: Cold Spring Harbor Laboratory Press.
Dale, R. et al. (1985). Plasmid. 13, 31-40.
Kunkel, T.A. et al. (1987). Methods Enzymol.. 154, 367-382.
Panet, A. et al. (1973). Biochemistry. 12, 5045-5050.
操作说明、说明书 & 用法
操作说明
Protocol for blunting ends by 3’ overhang removal and 3’ recessed (5’ overhang) end fill-in using T4 DNA Polymerase (M0203)
使用指南
Activity of DNA Modifying Enzymes in rCutSmart™ Buffer
工具 & 资源
选择指南
Blunting Selection Chart
Common Applications for Exonucleases and Endonucleases
DNA Polymerase Selection Chart
Web 工具
NEBcloner®
FAQs & 问题解决指南
FAQs
Can T4 DNA Polymerase be used in other NEBuffers and rCutSmart Buffer?
Can T4 DNA Polymerase be used to blunt DNA?
Can T4 DNA Polymerase be used to fill in 3′ overhangs?
Can T4 DNA Polymerase be used to remove 5′ overhangs?
Can T4 DNA Polymerase be heat inactivated?
Are the nucleotides needed to remove a 3′ overhang using T4 DNA Polymerase?
What are the main causes of blunting reaction failure using T4 DNA Polymerase?
Can T4 DNA Polymerase be used in labeling reactions and partial fill in reactions?
Is T4 DNA Polymerase active at room temperature?
Is T4 DNA Polymerase the enzyme of choice for removing 3′ overhangs and filling in 5′ overhangs (3′ recessed ends)?
Are NEB DNA Polymerases supplied with dNTPs?
Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
实验技巧
Excess enzyme, temperatures above 12°C, or limited dNTP concentrations can cause excessive 3’ → 5’ exonuclease degradation, which eliminates blunt end formation.
上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。
K. lactis Protein Expression Kit
The K. lactis Protein Expression Kit provides an easy method for expressing a gene of interest in the yeast Kluyveromyces lactis. Proteins may be produced intracellularly or secreted using an integrative expression vector (pKLAC2). To achieve protein secretion, a gene of interest is cloned downstream of the K. lactis α-mating factor secretion domain, which is eventually processed in the Golgi and results in secretion of the desired protein.
Supplied with competent K. lactis cells (NEB# C1001)
Allows cloning and expression of genes toxic to E. coli
Rapid high cell density growth results in high yield protein expression
Easy-to-use protocols for those inexperienced with yeast systems
上海金畔生物科技有限公司代理New England Biolabs(NEB)酶试剂全线产品,欢迎访问官网了解更多产品信息和订购。
EcoGII Methyltransferase
We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.
EcoGII Methyltransferase is a non-specific methyltransferase that modifies adenine residues (N6) in any sequence context.
Supplied with 10x rCutSmart™ Buffer and 32 mM S-adenosylmethionine (SAM)
This is an Enzyme for Innovation (EFI). EFI is a project initiated by NEB to provide unique enzymes to the scientific community in the hopes of enabling the discovery of new and innovative applications. These enzymes have interesting properties and unique specificities.