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
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
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)在不同缓冲液中的活性
NEBuffer™ 1: 60%
NEBuffer™ 2: 100%
NEBuffer™ 3: 100%
NEBuffer™ 4: 100%贮存溶液
100 mM KPO4
1 mM DTT
50% Glycerol
pH 6.5 @ 25°C热失活
75°C for 20 min
分子量
理论上的: 104000 daltons
5′ – 3′ 核酸外切酶
No
3′ – 5′ 核酸外切酶
Yes
链置换
No
单位活性检测条件
1X NEBuffer 2.1, 33 µM dNTPs including [3H]-dTTP, 70 µg/ml denatured herring sperm DNA.
错配率
~ 1×10-6bases
- 优势和特性
应用特性
- 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).
- 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.
引用 & 技术文献
- 引用文献
产品引用文献查找工具
Powered by Bioz See more details on Bioz
质控、安全 & 法规
- 质控分析
每一新批次的 NEB 产品都要经过质控，以满足指定的质量标准，对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息，可从此处查阅。
- 产品说明与变更通知
产品说明与变更通知
产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]
- M0203S_L_v1
- CoA
CoA 文件包含单个批次产品的储存温度、有效期和质控，这些文件的命名规则如下： [货号]_[规格]_[版本]_[Lot#]
- 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
- SDS
以下 SDS 文件可以帮助您安全地使用该产品
-
T4 DNA Polymerase
-
NEBuffer™ r2.1
-
T4 DNA Polymerase |NEB酶试剂 New England Biolabs
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
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
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)在不同缓冲液中的活性
NEBuffer™ 1: 60%
NEBuffer™ 2: 100%
NEBuffer™ 3: 100%
NEBuffer™ 4: 100%贮存溶液
100 mM KPO4
1 mM DTT
50% Glycerol
pH 6.5 @ 25°C热失活
75°C for 20 min
分子量
理论上的: 104000 daltons
5′ – 3′ 核酸外切酶
No
3′ – 5′ 核酸外切酶
Yes
链置换
No
单位活性检测条件
1X NEBuffer 2.1, 33 µM dNTPs including [3H]-dTTP, 70 µg/ml denatured herring sperm DNA.
错配率
~ 1×10-6bases
- 优势和特性
应用特性
- 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).
- 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.
引用 & 技术文献
- 引用文献
产品引用文献查找工具
Powered by Bioz See more details on Bioz
质控、安全 & 法规
- 质控分析
每一新批次的 NEB 产品都要经过质控，以满足指定的质量标准，对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息，可从此处查阅。
- 产品说明与变更通知
产品说明与变更通知
产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]
- M0203S_L_v1
- CoA
CoA 文件包含单个批次产品的储存温度、有效期和质控，这些文件的命名规则如下： [货号]_[规格]_[版本]_[Lot#]
- 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
- SDS
以下 SDS 文件可以帮助您安全地使用该产品
-
T4 DNA Polymerase
-
NEBuffer™ r2.1
-
neb P0704L说明书
neb P0704L说明书
PNGase F.
目录 ＃
P0704S
-
浓度
500,000单位/毫升
尺寸
-
P0704L 75,000个单位
价格表
$ 718.00
PNGase F是从糖蛋白中去除几乎所有N-连接的寡糖的有效的酶促方法。PNGase F是酰胺酶，其在高甘露糖，杂合和复合寡糖的内部GlcNAc和天冬酰胺残基之间切割。
- 通过SDS-PAGE和完整的ESI-MS测定，纯度≥95％
- 非重组，没有可检测的内切糖苷酶F1，F2或F3污染
- 储存在50％甘油中; 宜活动和稳定性长达24个月
- 可在天然或变性条件下使用
- 优化糖蛋白的去糖基化; 使N-聚糖核心寡糖保持完整，适合进一步分析
肽 – N-糖苷酶F，也称为PNGase F，是一种酰胺酶，可裂解来自N-连接糖蛋白的高甘露糖，杂合寡糖和复合寡糖的内层GlcNAc和天冬酰胺残基（1）
详细的特异性：
当内部的GlcNAc残基与α1-3岩藻糖残基连接时，PNGase F不能从糖蛋白切割N-连接的聚糖。这种修饰常见于植物和一些昆虫糖蛋白中。
产品来源
PNGase F从脑膜炎黄杆菌（Flavobacterium meningosepticum）（3）中纯化，不含蛋白酶和Endo F活性。
提供的试剂
本产品随附以下试剂：
| 存储在（°C） | 浓度 | |
| 糖蛋白变性缓冲液 | -20 | 10 X. |
| NP-40 | -20 | 10％ |
| GlycoBuffer 2 | -20 | 10 X. |
应用功能
- 糖蛋白分析
- 从糖蛋白中去除高甘露糖，杂合和复杂的 N-聚糖
- 无污染物（Endo F，蛋白酶等）
单位定义
一个单位定义为在37℃下在10μl的总反应体积中在1小时内从10μg变性的RNase B中除去> 95％的碳水化合物所需的酶量。
单位定义测定：
将10μgRNaseB用1X糖蛋白变性缓冲液（0.5％SDS，40mM DTT）在100℃变性10分钟。加入NP-40和GlycoBuffer 2后，加入两倍稀释的PNGase F，将反应混合物在37℃下温育1小时。通过SDS-PAGE显现反应产物的分离。
1X糖蛋白变性缓冲液
0.5％SDS
40 mM DTT
1X NP-40
1％NP-40，MilliQ-H 2 O
反应条件
1X GlycoBuffer 2
在37°C孵育
1X GlycoBuffer 2
50 mM磷酸钠
（pH值@ 25°C）
贮存温度
-20℃下
储藏条件
在 25℃下，20mM Tris-HCl
50mM NaCl
5mM EDTA
50％甘油
pH 7.5
热灭活
75°C，10分钟
分子量
表观：36000道尔顿
- 由于SDS抑制PNGase F活性，因此必须使NP-40存在于反应混合物中。为什么这种非离子型去污剂抵消SDS抑制作用目前尚不清楚。
- 为了使天然糖蛋白去糖基化，可能需要更长的孵育时间以及更多的酶。
- PNGase F不会切割 含有核心α1-3岩藻糖的N-连接聚糖。
- 典型反应条件：请参阅协议
Bst DNA Polymerase, Large Fragment | NEB酶试剂 New England Biolabs
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。
Bst DNA Polymerase, Large Fragment
The original polymerase for Loop-Mediated Isothermal Amplification (LAMP)
- Modified to retain 5´ → 3´ polymerase activity while lacking 5´ →3´ exonuclease activity
- Suitable for applications requiring thermophilic strand displacement
- Supplied with ThermoPol® Reaction Buffer for high product yields in demanding conditions
Interested in glycerol-free or custom formulations? – contact our Custom Solutions Group
Bulk packaging may also be available and requested for large recurring orders.
Learn More
产品信息
Bst DNA Polymerase, Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the
5´ → 3´ polymerase activity, but lacks 5´ →3´ exonuclease activity.
重点
- Isolated from a recombinant source
- Sequencing through problematic secondary structures
- Supplied with 10X Reaction Buffer
产品来源
Bst Polymerase, Large Fragment is prepared from an E. coli strain containing a genetic fusion of the Bacillus stearothermophilus DNA Polymerase gene, lacking the 5´ → 3´ exonuclease domain, and the gene coding for E. coli maltose binding protein (MBP). The fusion protein is purified to near homogeneity and the MBP portion of the fusion is cleaved off in vitro. The remaining polymerase is purified free of MBP (1).
- 产品类别:
- Isothermal Amplification & Strand Displacement Products
- 应用:
- Whole Genome Amplification,
- Loop-Mediated Isothermal Amplification,
- Isothermal Amplification
- 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 -
M0275V -20 Bst DNA Polymerase, Large Fragment M0275VVIAL -20 1 x 0.1 ml 8,000 units/ml ThermoPol® Reaction Buffer Pack B9004SVIAL -20 1 x 1.5 ml 10 X Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
-
M0275S -20 Bst DNA Polymerase, Large Fragment M0275SVIAL -20 1 x 0.2 ml 8,000 units/ml ThermoPol® Reaction Buffer Pack B9004SVIAL -20 1 x 1.5 ml 10 X Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
-
M0275L -20 Bst DNA Polymerase, Large Fragment M0275LVIAL -20 1 x 1 ml 8,000 units/ml ThermoPol® Reaction Buffer Pack B9004SVIAL -20 2 x 1.5 ml 10 X Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
-
M0275M -20 Bst DNA Polymerase, Large Fragment M0275MVIAL -20 1 x 0.067 ml 120,000 units/ml ThermoPol® Reaction Buffer Pack B9004SVIAL -20 2 x 1.5 ml 10 X Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
-
- 特性和用法
单位定义
One unit is defined at the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.
反应条件
1X ThermoPol® Reaction Buffer Pack
Incubate at 65°C1X ThermoPol® Reaction Buffer Pack
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton® X-100
(pH 8.8 @ 25°C)在不同缓冲液中的活性
NEBuffer™ 1: 50%
NEBuffer™ 2: 100%
NEBuffer™ 3: 50%
NEBuffer™ 4: 100%贮存溶液
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.1% Triton® X-100
pH 7.1 @ 25°C热失活
80°C for 20 min
分子量
理论上的: 67000 daltons
5′ – 3′ 核酸外切酶
No
3′ – 5′ 核酸外切酶
No
链置换
++++
单位活性检测条件
50 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM MgCl2, 30 nM M13mp18 SS DNA, 70 nM M13 sequencing primer (–47) 24 mer, 200 µM dATP, 200 µM dCTP, 200 µM dGTP, 100 µM dTTP including [3H]-dTTP, and 100 µg/ml BSA.
- 优势和特性
应用特性
- Isothermal amplification (LAMP)
- DNA sequencing through high GC regions (2,3)
- Rapid Sequencing from nanogram amounts of DNA template (4)
- 相关产品
相关产品
- Deoxynucleotide (dNTP) Solution Set
- Magnesium Sulfate (MgSO4) Solution
- Deoxynucleotide Solution Mix
- Bst 2.0 DNA Polymerase
- Bst 2.0 WarmStart DNA Polymerase
单独销售的组分
- ThermoPol® Reaction Buffer Pack
- 注意事项
- Bst DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
- 100 µg/ml BSA or 0.1%Triton X-100 is required for long term storage.
- Reaction temperatures above 70°C are not recommended.
- Bst DNA Polymerase, Large Fragment cannot be used for thermal cycle sequencing or PCR.
- 参考文献
- Kong, H., Aliotta, J. and Pelletier, J.J. New England Biolabs. Unpublished observation
- Griffin, H. and Griffin, A. (1994). PCRTechnology. 228-229.
- McClary, J. et al. (1991). J. DNASequencing and Mapping. 1, 173-180.
- Mead, D.A. et al. (1991). Biotechniques. 11, 76-87.
- Notomi et al. (2000). Nucleic Acids Res. 28(12), E63.
- Hsieh et al. (2014). Chem. Commun. 50, 3747-3749.
- Tanner and Evans (2014). Curr. Prot. Mol. Biol. 105, 15.14.
操作说明、说明书 & 用法
- 操作说明
- Typical LAMP Protocol (M0275)
- 使用指南
- Activity of DNA Modifying Enzymes in rCutSmart™ Buffer
工具 & 资源
- 选择指南
- DNA Polymerase Selection Chart
- Web 工具
- NEB LAMP Primer Design Tool
FAQs & 问题解决指南
- FAQs
- Can Bst DNA Polymerase be used in other NEBuffers, including rCutSmart?
- Can Bst DNA Polymerase be used to blunt DNA?
- Can Bst DNA Polymerase be used to fill in 3′ overhangs?
- Can Bst DNA Polymerase be used to remove 5′ overhangs?
- Can Bst DNA Polymerase be heat inactivated?
- Are NEB DNA Polymerases supplied with dNTPs?
- What are the main causes of reaction failure using Bst DNA Polymerase?
- Does Bst DNA Polymerase have an active 3’→5′ proofreading exonuclease?
- Can Bst DNA Polymerase be used for thermal cycle sequencing?
- Can Bst DNA Polymerase initiate at a nick in the DNA?
- Can Bst DNA Polymerase be used in labeling reactions and partial fill in reactions?
- Can Bst DNA Polymerase be diluted?
- When should Bst DNA Polymerase be the enzyme of choice?
- Can Bst DNA Polymerase be used at temperatures other than 65°C?
- Does Bst DNA Polymerase, Large Fragment incorporate dUTP?
- Does Bst DNA polymerase have reverse transcriptase activity?
- Does NEB have a master mix for LAMP or RT-LAMP reactions?
- What is LAMP and RT-LAMP?
- 问题解决指南
- PCR Troubleshooting Guide
引用 & 技术文献
- 引用文献
产品引用文献查找工具
Powered by Bioz See more details on Bioz更多引用文献
- Trisadee Khamlor, Petai Pongpiachan, Rangsun Parnpai, Kanchana Punyawai, Siwat Sangsritavong, Nipa Chokesajjawatee (2015) Bovine embryo sex determination by multiplex loop-mediated isothermal amplification. Theriogenology; 83, 891-6. PubMedID: 25542460, DOI: 10.1016/j.theriogenology.2014.11.025
- DoKyung Lee, Eun Jin Kim, Paul E Kilgore, Soon Ae Kim, Hideyuki Takahashi, Makoto Ohnishi, Dang Duc Anh, Bai Qing Dong, Jung Soo Kim, Jun Tomono, Shigehiko Miyamoto, Tsugunori Notomi, Dong Wook Kim, Mitsuko Seki (2015) Clinical Evaluation of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Neisseria meningitidis in Cerebrospinal Fluid. PLoS One; 10, e0122922. PubMedID: 25853422, DOI: 10.1371/journal.pone.0122922
- Manabu Nemoto, Yoshinori Morita, Hidekazu Niwa, Hiroshi Bannai, Koji Tsujimura, Takashi Yamanaka, Takashi Kondo (2015) Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification. J Virol Methods; 215-216, 13-6. PubMedID: 25682750, DOI: 10.1016/j.jviromet.2015.02.001
- Mohammad Reza Allahyar Torkaman, Kazunari Kamachi, Vajihe Sadat Nikbin, Masoumeh Nakhost Lotfi, Fereshteh Shahcheraghi (2015) Comparison of loop-mediated isothermal amplification and real-time PCR for detecting Bordetella pertussis. J Med Microbiol; 64, 463-5. PubMedID: 25596118, DOI: 10.1099/jmm.0.000021
- Aongart Mahittikorn, Hirotake Mori, Supaluk Popruk, Amonrattana Roobthaisong, Chantira Sutthikornchai, Khuanchai Koompapong, Sukhontha Siri, Yaowalark Sukthana, Duangporn Nacapunchai (2015) Development of a Rapid, Simple Method for Detecting Naegleria fowleri Visually in Water Samples by Loop-Mediated Isothermal Amplification (LAMP). PLoS One; 10, e0120997. PubMedID: 25822175, DOI: 10.1371/journal.pone.0120997
质控、安全 & 法规
- 质控分析
每一新批次的 NEB 产品都要经过质控，以满足指定的质量标准，对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息，可从此处查阅。
- 产品说明与变更通知
产品说明与变更通知
产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]
- M0275M_v1
- M0275S_L_v1
- M0275M_v2
- M0275S_L_v2
- CoA
CoA 文件包含单个批次产品的储存温度、有效期和质控，这些文件的命名规则如下： [货号]_[规格]_[版本]_[Lot#]
- M0275S_L_v1_0511509
- M0275M_v1_0511509
- M0275M_v1_0511603
- M0275S_L_v1_0511603
- M0275M_v1_0511609
- M0275S_L_v1_0511609
- M0275S_L_v1_0511703
- M0275M_v1_0511703
- M0275S_L_v1_0511709
- M0275M_v1_0511709
- M0275M_v1_0511803
- M0275S_L_v1_0511803
- M0275L_v1_10011610
- M0275S_v1_10009342
- M0275M_v1_10017700
- M0275L_v1_10020437
- M0275S_v1_10032197
- M0275L_v1_10029594
- M0275S_v1_10038935
- M0275M_v1_10035567
- M0275L_v1_10046214
- M0275S_v1_10045900
- M0275S_v1_10048185
- M0275L_v1_10051813
- M0275M_v1_10055954
- M0275L_v1_10053202
- M0275S_v1_10059774
- M0275S_v2_10063500
- M0275L_v1_10063499
- M0275L_v1_10067894
- M0275M_v2_10068169
- M0275S_v2_10068243
- M0275L_v2_10072178
- M0275M_v2_10073620
- M0275S_v2_10078286
- M0275L_v2_10075398
- M0275M_v2_10086262
- M0275S_v2_10088116
- M0275S_v2_10094232
- M0275M_v2_10095463
- M0275L_v2_10100585
- M0275S_v2_10108804
- M0275S_v2_10115484
- M0275L_v2_10108569
- M0275M_v2_10127396
- M0275S_v2_10122371
- M0275L_v2_10136893
- M0275M_v2_10140685
- M0275S_v2_10153544
- M0275M_v2_10160590
- M0275M_v2_10164128
- M0275L_v2_10162366
- SDS
以下 SDS 文件可以帮助您安全地使用该产品
-
Bst DNA Polymerase, Large Fragment
-
ThermoPol® Reaction Buffer Pack
-
Magnesium Sulfate (MgSO4) Solution
-
Bst DNA Polymerase, Large Fragment |NEB酶试剂 New England Biolabs
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。
Bst DNA Polymerase, Large Fragment
The original polymerase for Loop-Mediated Isothermal Amplification (LAMP)
- Modified to retain 5´ → 3´ polymerase activity while lacking 5´ →3´ exonuclease activity
- Suitable for applications requiring thermophilic strand displacement
- Supplied with ThermoPol® Reaction Buffer for high product yields in demanding conditions
Interested in glycerol-free or custom formulations? – contact our Custom Solutions Group
Bulk packaging may also be available and requested for large recurring orders.
Learn More
产品信息
Bst DNA Polymerase, Large Fragment is the portion of the Bacillus stearothermophilus DNA Polymerase protein that contains the
5´ → 3´ polymerase activity, but lacks 5´ →3´ exonuclease activity.
重点
- Isolated from a recombinant source
- Sequencing through problematic secondary structures
- Supplied with 10X Reaction Buffer
产品来源
Bst Polymerase, Large Fragment is prepared from an E. coli strain containing a genetic fusion of the Bacillus stearothermophilus DNA Polymerase gene, lacking the 5´ → 3´ exonuclease domain, and the gene coding for E. coli maltose binding protein (MBP). The fusion protein is purified to near homogeneity and the MBP portion of the fusion is cleaved off in vitro. The remaining polymerase is purified free of MBP (1).
- 产品类别:
- Isothermal Amplification & Strand Displacement Products
- 应用:
- Whole Genome Amplification,
- Loop-Mediated Isothermal Amplification,
- Isothermal Amplification
- 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 -
M0275V -20 Bst DNA Polymerase, Large Fragment M0275VVIAL -20 1 x 0.1 ml 8,000 units/ml ThermoPol® Reaction Buffer Pack B9004SVIAL -20 1 x 1.5 ml 10 X Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
-
M0275S -20 Bst DNA Polymerase, Large Fragment M0275SVIAL -20 1 x 0.2 ml 8,000 units/ml ThermoPol® Reaction Buffer Pack B9004SVIAL -20 1 x 1.5 ml 10 X Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
-
M0275L -20 Bst DNA Polymerase, Large Fragment M0275LVIAL -20 1 x 1 ml 8,000 units/ml ThermoPol® Reaction Buffer Pack B9004SVIAL -20 2 x 1.5 ml 10 X Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
-
M0275M -20 Bst DNA Polymerase, Large Fragment M0275MVIAL -20 1 x 0.067 ml 120,000 units/ml ThermoPol® Reaction Buffer Pack B9004SVIAL -20 2 x 1.5 ml 10 X Magnesium Sulfate (MgSO4) Solution B1003SVIAL -20 1 x 1.5 ml 100 mM
-
- 特性和用法
单位定义
One unit is defined at the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 65°C.
反应条件
1X ThermoPol® Reaction Buffer Pack
Incubate at 65°C1X ThermoPol® Reaction Buffer Pack
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton® X-100
(pH 8.8 @ 25°C)在不同缓冲液中的活性
NEBuffer™ 1: 50%
NEBuffer™ 2: 100%
NEBuffer™ 3: 50%
NEBuffer™ 4: 100%贮存溶液
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
0.1% Triton® X-100
pH 7.1 @ 25°C热失活
80°C for 20 min
分子量
理论上的: 67000 daltons
5′ – 3′ 核酸外切酶
No
3′ – 5′ 核酸外切酶
No
链置换
++++
单位活性检测条件
50 mM KCl, 20 mM Tris-HCl (pH 8.8), 10 mM MgCl2, 30 nM M13mp18 SS DNA, 70 nM M13 sequencing primer (–47) 24 mer, 200 µM dATP, 200 µM dCTP, 200 µM dGTP, 100 µM dTTP including [3H]-dTTP, and 100 µg/ml BSA.
- 优势和特性
应用特性
- Isothermal amplification (LAMP)
- DNA sequencing through high GC regions (2,3)
- Rapid Sequencing from nanogram amounts of DNA template (4)
- 相关产品
相关产品
- Deoxynucleotide (dNTP) Solution Set
- Magnesium Sulfate (MgSO4) Solution
- Deoxynucleotide Solution Mix
- Bst 2.0 DNA Polymerase
- Bst 2.0 WarmStart DNA Polymerase
单独销售的组分
- ThermoPol® Reaction Buffer Pack
- 注意事项
- Bst DNA Polymerase does not exhibit 3´→ 5´ exonuclease activity.
- 100 µg/ml BSA or 0.1%Triton X-100 is required for long term storage.
- Reaction temperatures above 70°C are not recommended.
- Bst DNA Polymerase, Large Fragment cannot be used for thermal cycle sequencing or PCR.
- 参考文献
- Kong, H., Aliotta, J. and Pelletier, J.J. New England Biolabs. Unpublished observation
- Griffin, H. and Griffin, A. (1994). PCRTechnology. 228-229.
- McClary, J. et al. (1991). J. DNASequencing and Mapping. 1, 173-180.
- Mead, D.A. et al. (1991). Biotechniques. 11, 76-87.
- Notomi et al. (2000). Nucleic Acids Res. 28(12), E63.
- Hsieh et al. (2014). Chem. Commun. 50, 3747-3749.
- Tanner and Evans (2014). Curr. Prot. Mol. Biol. 105, 15.14.
操作说明、说明书 & 用法
- 操作说明
- Typical LAMP Protocol (M0275)
- 使用指南
- Activity of DNA Modifying Enzymes in rCutSmart™ Buffer
工具 & 资源
- 选择指南
- DNA Polymerase Selection Chart
- Web 工具
- NEB LAMP Primer Design Tool
FAQs & 问题解决指南
- FAQs
- Can Bst DNA Polymerase be used in other NEBuffers, including rCutSmart?
- Can Bst DNA Polymerase be used to blunt DNA?
- Can Bst DNA Polymerase be used to fill in 3′ overhangs?
- Can Bst DNA Polymerase be used to remove 5′ overhangs?
- Can Bst DNA Polymerase be heat inactivated?
- Are NEB DNA Polymerases supplied with dNTPs?
- What are the main causes of reaction failure using Bst DNA Polymerase?
- Does Bst DNA Polymerase have an active 3’→5′ proofreading exonuclease?
- Can Bst DNA Polymerase be used for thermal cycle sequencing?
- Can Bst DNA Polymerase initiate at a nick in the DNA?
- Can Bst DNA Polymerase be used in labeling reactions and partial fill in reactions?
- Can Bst DNA Polymerase be diluted?
- When should Bst DNA Polymerase be the enzyme of choice?
- Can Bst DNA Polymerase be used at temperatures other than 65°C?
- Does Bst DNA Polymerase, Large Fragment incorporate dUTP?
- Does Bst DNA polymerase have reverse transcriptase activity?
- Does NEB have a master mix for LAMP or RT-LAMP reactions?
- What is LAMP and RT-LAMP?
- 问题解决指南
- PCR Troubleshooting Guide
引用 & 技术文献
- 引用文献
产品引用文献查找工具
Powered by Bioz See more details on Bioz更多引用文献
- Trisadee Khamlor, Petai Pongpiachan, Rangsun Parnpai, Kanchana Punyawai, Siwat Sangsritavong, Nipa Chokesajjawatee (2015) Bovine embryo sex determination by multiplex loop-mediated isothermal amplification. Theriogenology; 83, 891-6. PubMedID: 25542460, DOI: 10.1016/j.theriogenology.2014.11.025
- DoKyung Lee, Eun Jin Kim, Paul E Kilgore, Soon Ae Kim, Hideyuki Takahashi, Makoto Ohnishi, Dang Duc Anh, Bai Qing Dong, Jung Soo Kim, Jun Tomono, Shigehiko Miyamoto, Tsugunori Notomi, Dong Wook Kim, Mitsuko Seki (2015) Clinical Evaluation of a Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Neisseria meningitidis in Cerebrospinal Fluid. PLoS One; 10, e0122922. PubMedID: 25853422, DOI: 10.1371/journal.pone.0122922
- Manabu Nemoto, Yoshinori Morita, Hidekazu Niwa, Hiroshi Bannai, Koji Tsujimura, Takashi Yamanaka, Takashi Kondo (2015) Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification. J Virol Methods; 215-216, 13-6. PubMedID: 25682750, DOI: 10.1016/j.jviromet.2015.02.001
- Mohammad Reza Allahyar Torkaman, Kazunari Kamachi, Vajihe Sadat Nikbin, Masoumeh Nakhost Lotfi, Fereshteh Shahcheraghi (2015) Comparison of loop-mediated isothermal amplification and real-time PCR for detecting Bordetella pertussis. J Med Microbiol; 64, 463-5. PubMedID: 25596118, DOI: 10.1099/jmm.0.000021
- Aongart Mahittikorn, Hirotake Mori, Supaluk Popruk, Amonrattana Roobthaisong, Chantira Sutthikornchai, Khuanchai Koompapong, Sukhontha Siri, Yaowalark Sukthana, Duangporn Nacapunchai (2015) Development of a Rapid, Simple Method for Detecting Naegleria fowleri Visually in Water Samples by Loop-Mediated Isothermal Amplification (LAMP). PLoS One; 10, e0120997. PubMedID: 25822175, DOI: 10.1371/journal.pone.0120997
质控、安全 & 法规
- 质控分析
每一新批次的 NEB 产品都要经过质控，以满足指定的质量标准，对特定产品的质量标准和每一批次的检测数据可以通过产品说明表格、COA、产品信息卡或者产品手册进行查阅或下载。关于 NEB 产品质控的详细信息，可从此处查阅。
- 产品说明与变更通知
产品说明与变更通知
产品说明表包含产品的储存温度、有效期和规格，这些文件的命名规则如下：[货号]_[规格]_[版本]
- M0275M_v1
- M0275S_L_v1
- M0275M_v2
- M0275S_L_v2
- CoA
CoA 文件包含单个批次产品的储存温度、有效期和质控，这些文件的命名规则如下： [货号]_[规格]_[版本]_[Lot#]
- M0275S_L_v1_0511509
- M0275M_v1_0511509
- M0275M_v1_0511603
- M0275S_L_v1_0511603
- M0275M_v1_0511609
- M0275S_L_v1_0511609
- M0275S_L_v1_0511703
- M0275M_v1_0511703
- M0275S_L_v1_0511709
- M0275M_v1_0511709
- M0275M_v1_0511803
- M0275S_L_v1_0511803
- M0275L_v1_10011610
- M0275S_v1_10009342
- M0275M_v1_10017700
- M0275L_v1_10020437
- M0275S_v1_10032197
- M0275L_v1_10029594
- M0275S_v1_10038935
- M0275M_v1_10035567
- M0275L_v1_10046214
- M0275S_v1_10045900
- M0275S_v1_10048185
- M0275L_v1_10051813
- M0275M_v1_10055954
- M0275L_v1_10053202
- M0275S_v1_10059774
- M0275S_v2_10063500
- M0275L_v1_10063499
- M0275L_v1_10067894
- M0275M_v2_10068169
- M0275S_v2_10068243
- M0275L_v2_10072178
- M0275M_v2_10073620
- M0275S_v2_10078286
- M0275L_v2_10075398
- M0275M_v2_10086262
- M0275S_v2_10088116
- M0275S_v2_10094232
- M0275M_v2_10095463
- M0275L_v2_10100585
- M0275S_v2_10108804
- M0275S_v2_10115484
- M0275L_v2_10108569
- M0275M_v2_10127396
- M0275S_v2_10122371
- M0275L_v2_10136893
- M0275M_v2_10140685
- M0275S_v2_10153544
- M0275M_v2_10160590
- M0275M_v2_10164128
- M0275L_v2_10162366
- SDS
以下 SDS 文件可以帮助您安全地使用该产品
-
Bst DNA Polymerase, Large Fragment
-
ThermoPol® Reaction Buffer Pack
-
Magnesium Sulfate (MgSO4) Solution
-