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。
DNA Polymerase I, Large (Klenow) Fragment
DNA Polymerase I, Large (Klenow) fragment was originally derived as a proteolytic product of E.coli DNA polymerase that retains polymerase and 3’ —> 5’ exonuclease activity
- Removal of 3’ overhangs or fill-in of 5’ overhangs to form blunt ends
- Lacks 5’ —> 3’ exonuclease activity
- Generates probes using random primers
- Second strand cDNA synthesis
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
重点
产品来源
An E. coli strain that contains the E. coli polA gene that has had its 5’→3′ exonuclease domain removed.
- 产品类别:
- DNA Manipulation Products
- 应用:
- Blunting,
- DNA Sequencing,
- Polymerases for DNA Manipulation,
- RT-PCR & cDNA Synthesis,
- RT-qPCR, RT-PCR and cDNA Synthesis,
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 -
M0210V -20 DNA Polymerase I, Large (Klenow) Fragment M0210VVIAL -20 1 x 0.02 ml 5,000 units/ml NEBuffer™ 2 B7002SVIAL -20 1 x 1.25 ml 10 X
-
M0210S -20 DNA Polymerase I, Large (Klenow) Fragment M0210SVIAL -20 1 x 0.04 ml 5,000 units/ml NEBuffer™ 2 B7002SVIAL -20 1 x 1.25 ml 10 X
-
M0210L -20 DNA Polymerase I, Large (Klenow) Fragment M0210LVIAL -20 1 x 0.2 ml 5,000 units/ml NEBuffer™ 2 B7002SVIAL -20 1 x 1.25 ml 10 X
-
M0210M -20 DNA Polymerase I, Large (Klenow) Fragment M0210MVIAL -20 1 x 0.02 ml 50,000 units/ml NEBuffer™ 2 B7002SVIAL -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.
反应条件
1X NEBuffer™ 2
Incubate at 25°C1X NEBuffer™ 2
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
1 mM DTT
(pH 7.9 @ 25°C)贮存溶液
25 mM Tris-HCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C热失活
75°C for 20 min
分子量
理论上的: 68000 daltons
5′ – 3′ 核酸外切酶
No
3′ – 5′ 核酸外切酶
Yes
链置换
+
单位活性检测条件
1X NEBuffer 2, 33 μM dNTPs including [3H]-dTTP and 70 μg/ml denatured herring sperm DNA.
错配率
~ 18×10-6bases
- 优势和特性
应用特性
- DNA sequencing by the Sanger dideoxy method (2)
- Fill-in of 5´ overhangs to form blunt ends (3)
- Removal of 3´ overhangs to form blunt ends (3)
- Second strand cDNA synthesis
- Second strand synthesis in mutagenesis protocols (4)
- 相关产品
相关产品
- Deoxynucleotide (dNTP) Solution Set
- Deoxynucleotide (dNTP) Solution Mix
- Monarch® Plasmid Miniprep Kit
- Monarch® DNA Gel Extraction Kit
- Monarch® PCR & DNA Cleanup Kit (5 μg)
单独销售的组分
- NEBuffer 2
- 注意事项
- Protocol for blunting ends by 3′ overhang removal and 3′ recessed end fill-in:
- DNA should be dissolved in 1X NEBuffer 1-4 or T4 DNA Ligase Reaction Buffer supplemented with 33 μM each dNTP. Add 1 unit DNA Polymerase I, Large (Klenow) Fragment per microgram DNA and incubate 15 minutes at 25°C. Stop reaction by adding EDTA to a final concentration of 10 mM and heating at 75°C for 20 minutes.
- CAUTION: 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.
- When DNA Polymerase I, Large (Klenow) Fragment is used to sequence DNA using the dideoxy method of Sanger et al., 1 unit/5 μl reaction volume is recommended.
- DNA Polymerase I, Large (Klenow) Fragment is also active in all four NEBuffers and T4 DNA Ligase Reaction Buffer when supplemented with dNTPs.
- Protocol for blunting ends by 3′ overhang removal and 3′ recessed end fill-in:
- 参考文献
- Jacobsen, H., Klenow, H. and Overgaard-Hansen, K. (1974). Eur. J. Biochem.. 45, 623-627.
- Sanger, F. et al. (1977). Proc. Natl. Acad. Sci. USA. 74, 5463-5467.
- Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.), 5.40-5.43. Cold Spring Harbor: Cold Spring Harbor Laboratory Press.
- Gubler, U. (1987). In S.L. Berger and A.R. Kimmel(Ed.), Methods in Enzymology. 152, 330-335. San Diego: Academic Press.
- Bebenek, K., Joyce, C.M., Fitzgerald, M.P. and Kunkel, T.A. (1990). J. Bio. Chem . 265, 13878-13887.
操作说明、说明书 & 用法
- 操作说明
- Protocol for blunting ends by 3′ overhang removal and fill-in of 3′ recessed (5′ overhang) ends using DNA Polymerase I, Large (Klenow) Fragment (M0210)
- 使用指南
- Activity of DNA Modifying Enzymes in rCutSmart™ Buffer
工具 & 资源
- 选择指南
- Blunting Selection Chart
- DNA Polymerase Selection Chart
- Web 工具
- NEBcloner®
FAQs & 问题解决指南
- FAQs
- Can DNA Polymerase I, Large (Klenow) Fragment be used in other NEBuffers, including rCutSmart?
- Can DNA Polymerase I, Large (Klenow) Fragment be used to blunt DNA?
- Can DNA Polymerase I, Large (Klenow) Fragment be used to fill in 3′ overhangs?
- Can DNA Polymerase I, Large (Klenow) Fragment be used to chew back 5′ overhangs?
- Can DNA Polymerase I, Large (Klenow) Fragment be heat inactivated?
- Are the nucleotides needed to remove a 3′ overhang with DNA Polymerase I, Large (Klenow) Fragment?
- What are the main causes for blunting reaction failure using DNA Polymerase I, Large (Klenow) Fragment?
- Can DNA Polymerase I, Large (Klenow) Fragment be used in labeling reactions and partial fill in reactions?
- Is DNA Polymerase I, Large (Klenow) Fragment the enzyme of choice for chewing back 3′ overhangs and filling in 5′ overhangs (3′ recessed ends)?
- Are NEB DNA Polymerases supplied with dNTPs?
- 实验技巧
- Excess enzyme, temperatures above 25°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]
- M0210M_v1
- M0210S_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#]
- M0210M_v1_0891509
- M0210S_L_v1_0891509
- M0210S_L_v1_0891512
- M0210S_L_v1_0891603
- M0210M_v1_0891603
- M0210M_v1_0891608
- M0210S_L_v1_0891608
- M0210M_v1_0891703
- M0210S_L_v1_0891703
- M0210M_v1_0891709
- M0210S_L_v1_0891709
- M0210M_v1_0891803
- M0210S_L_v1_0891803
- M0210L_v1_10007540
- M0210L_v1_10025743
- M0210S_v1_10024368
- M0210L_v1_10028508
- M0210M_v1_10028233
- M0210S_v1_10028423
- M0210L_v1_10034214
- M0210M_v1_10034564
- M0210S_v1_10034001
- M0210L_v1_10036837
- M0210S_v1_10037080
- M0210L_v0_10041498
- M0210L_v1_10046211
- M0210S_v1_10043965
- M0210S_v1_10050337
- M0210L_v1_10053482
- M0210S_v1_10055962
- M0210L_v1_10057607
- M0210L_v1_10058269
- M0210S_v1_10058172
- M0210L_v1_10062077
- M0210M_v1_10057608
- M0210S_v1_10062081
- M0210L_v1_10065299
- M0210S_v1_10066373
- M0210L_v1_10075466
- M0210S_v1_10075464
- M0210S_v1_10079480
- M0210M_v1_10087408
- M0210L_v1_10088146
- M0210S_v1_10087380
- M0210S_v1_10094981
- M0210L_v1_10100634
- M0210M_v1_10101977
- M0210L_v1_10106852
- M0210S_v1_10107645
- M0210S_v1_10113999
- M0210L_v1_10117205
- M0210M_v1_10117349
- M0210M_v1_10123077
- M0210S_v1_10127909
- M0210L_v1_10136292
- M0210M_v1_10137266
- M0210L_v1_10149552
- M0210S_v1_10150626
- M0210L_v1_10155130
- M0210M_v1_10154966
- M0210L_v1_10162372
- SDS
以下 SDS 文件可以帮助您安全地使用该产品
-
DNA Polymerase I, Large (Klenow) Fragment
-
NEBuffer™ 2
-
DNA Polymerase I, Large (Klenow) 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。
DNA Polymerase I, Large (Klenow) Fragment
DNA Polymerase I, Large (Klenow) fragment was originally derived as a proteolytic product of E.coli DNA polymerase that retains polymerase and 3’ —> 5’ exonuclease activity
- Removal of 3’ overhangs or fill-in of 5’ overhangs to form blunt ends
- Lacks 5’ —> 3’ exonuclease activity
- Generates probes using random primers
- Second strand cDNA synthesis
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
重点
产品来源
An E. coli strain that contains the E. coli polA gene that has had its 5’→3′ exonuclease domain removed.
- 产品类别:
- DNA Manipulation Products
- 应用:
- Blunting,
- DNA Sequencing,
- Polymerases for DNA Manipulation,
- RT-PCR & cDNA Synthesis,
- RT-qPCR, RT-PCR and cDNA Synthesis,
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 -
M0210V -20 DNA Polymerase I, Large (Klenow) Fragment M0210VVIAL -20 1 x 0.02 ml 5,000 units/ml NEBuffer™ 2 B7002SVIAL -20 1 x 1.25 ml 10 X
-
M0210S -20 DNA Polymerase I, Large (Klenow) Fragment M0210SVIAL -20 1 x 0.04 ml 5,000 units/ml NEBuffer™ 2 B7002SVIAL -20 1 x 1.25 ml 10 X
-
M0210L -20 DNA Polymerase I, Large (Klenow) Fragment M0210LVIAL -20 1 x 0.2 ml 5,000 units/ml NEBuffer™ 2 B7002SVIAL -20 1 x 1.25 ml 10 X
-
M0210M -20 DNA Polymerase I, Large (Klenow) Fragment M0210MVIAL -20 1 x 0.02 ml 50,000 units/ml NEBuffer™ 2 B7002SVIAL -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.
反应条件
1X NEBuffer™ 2
Incubate at 25°C1X NEBuffer™ 2
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
1 mM DTT
(pH 7.9 @ 25°C)贮存溶液
25 mM Tris-HCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C热失活
75°C for 20 min
分子量
理论上的: 68000 daltons
5′ – 3′ 核酸外切酶
No
3′ – 5′ 核酸外切酶
Yes
链置换
+
单位活性检测条件
1X NEBuffer 2, 33 μM dNTPs including [3H]-dTTP and 70 μg/ml denatured herring sperm DNA.
错配率
~ 18×10-6bases
- 优势和特性
应用特性
- DNA sequencing by the Sanger dideoxy method (2)
- Fill-in of 5´ overhangs to form blunt ends (3)
- Removal of 3´ overhangs to form blunt ends (3)
- Second strand cDNA synthesis
- Second strand synthesis in mutagenesis protocols (4)
- 相关产品
相关产品
- Deoxynucleotide (dNTP) Solution Set
- Deoxynucleotide (dNTP) Solution Mix
- Monarch® Plasmid Miniprep Kit
- Monarch® DNA Gel Extraction Kit
- Monarch® PCR & DNA Cleanup Kit (5 μg)
单独销售的组分
- NEBuffer 2
- 注意事项
- Protocol for blunting ends by 3′ overhang removal and 3′ recessed end fill-in:
- DNA should be dissolved in 1X NEBuffer 1-4 or T4 DNA Ligase Reaction Buffer supplemented with 33 μM each dNTP. Add 1 unit DNA Polymerase I, Large (Klenow) Fragment per microgram DNA and incubate 15 minutes at 25°C. Stop reaction by adding EDTA to a final concentration of 10 mM and heating at 75°C for 20 minutes.
- CAUTION: 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.
- When DNA Polymerase I, Large (Klenow) Fragment is used to sequence DNA using the dideoxy method of Sanger et al., 1 unit/5 μl reaction volume is recommended.
- DNA Polymerase I, Large (Klenow) Fragment is also active in all four NEBuffers and T4 DNA Ligase Reaction Buffer when supplemented with dNTPs.
- Protocol for blunting ends by 3′ overhang removal and 3′ recessed end fill-in:
- 参考文献
- Jacobsen, H., Klenow, H. and Overgaard-Hansen, K. (1974). Eur. J. Biochem.. 45, 623-627.
- Sanger, F. et al. (1977). Proc. Natl. Acad. Sci. USA. 74, 5463-5467.
- Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual. (2nd ed.), 5.40-5.43. Cold Spring Harbor: Cold Spring Harbor Laboratory Press.
- Gubler, U. (1987). In S.L. Berger and A.R. Kimmel(Ed.), Methods in Enzymology. 152, 330-335. San Diego: Academic Press.
- Bebenek, K., Joyce, C.M., Fitzgerald, M.P. and Kunkel, T.A. (1990). J. Bio. Chem . 265, 13878-13887.
操作说明、说明书 & 用法
- 操作说明
- Protocol for blunting ends by 3′ overhang removal and fill-in of 3′ recessed (5′ overhang) ends using DNA Polymerase I, Large (Klenow) Fragment (M0210)
- 使用指南
- Activity of DNA Modifying Enzymes in rCutSmart™ Buffer
工具 & 资源
- 选择指南
- Blunting Selection Chart
- DNA Polymerase Selection Chart
- Web 工具
- NEBcloner®
FAQs & 问题解决指南
- FAQs
- Can DNA Polymerase I, Large (Klenow) Fragment be used in other NEBuffers, including rCutSmart?
- Can DNA Polymerase I, Large (Klenow) Fragment be used to blunt DNA?
- Can DNA Polymerase I, Large (Klenow) Fragment be used to fill in 3′ overhangs?
- Can DNA Polymerase I, Large (Klenow) Fragment be used to chew back 5′ overhangs?
- Can DNA Polymerase I, Large (Klenow) Fragment be heat inactivated?
- Are the nucleotides needed to remove a 3′ overhang with DNA Polymerase I, Large (Klenow) Fragment?
- What are the main causes for blunting reaction failure using DNA Polymerase I, Large (Klenow) Fragment?
- Can DNA Polymerase I, Large (Klenow) Fragment be used in labeling reactions and partial fill in reactions?
- Is DNA Polymerase I, Large (Klenow) Fragment the enzyme of choice for chewing back 3′ overhangs and filling in 5′ overhangs (3′ recessed ends)?
- Are NEB DNA Polymerases supplied with dNTPs?
- 实验技巧
- Excess enzyme, temperatures above 25°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]
- M0210M_v1
- M0210S_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#]
- M0210M_v1_0891509
- M0210S_L_v1_0891509
- M0210S_L_v1_0891512
- M0210S_L_v1_0891603
- M0210M_v1_0891603
- M0210M_v1_0891608
- M0210S_L_v1_0891608
- M0210M_v1_0891703
- M0210S_L_v1_0891703
- M0210M_v1_0891709
- M0210S_L_v1_0891709
- M0210M_v1_0891803
- M0210S_L_v1_0891803
- M0210L_v1_10007540
- M0210L_v1_10025743
- M0210S_v1_10024368
- M0210L_v1_10028508
- M0210M_v1_10028233
- M0210S_v1_10028423
- M0210L_v1_10034214
- M0210M_v1_10034564
- M0210S_v1_10034001
- M0210L_v1_10036837
- M0210S_v1_10037080
- M0210L_v0_10041498
- M0210L_v1_10046211
- M0210S_v1_10043965
- M0210S_v1_10050337
- M0210L_v1_10053482
- M0210S_v1_10055962
- M0210L_v1_10057607
- M0210L_v1_10058269
- M0210S_v1_10058172
- M0210L_v1_10062077
- M0210M_v1_10057608
- M0210S_v1_10062081
- M0210L_v1_10065299
- M0210S_v1_10066373
- M0210L_v1_10075466
- M0210S_v1_10075464
- M0210S_v1_10079480
- M0210M_v1_10087408
- M0210L_v1_10088146
- M0210S_v1_10087380
- M0210S_v1_10094981
- M0210L_v1_10100634
- M0210M_v1_10101977
- M0210L_v1_10106852
- M0210S_v1_10107645
- M0210S_v1_10113999
- M0210L_v1_10117205
- M0210M_v1_10117349
- M0210M_v1_10123077
- M0210S_v1_10127909
- M0210L_v1_10136292
- M0210M_v1_10137266
- M0210L_v1_10149552
- M0210S_v1_10150626
- M0210L_v1_10155130
- M0210M_v1_10154966
- M0210L_v1_10162372
- SDS
以下 SDS 文件可以帮助您安全地使用该产品
-
DNA Polymerase I, Large (Klenow) Fragment
-
NEBuffer™ 2
-
X-Large Cell and Tissue Collection Kits
X-Large Cell and Tissue Collection Kits
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详细介绍
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X-Large Cell and Tissue Collection Kits
CTK40-X适用于常规组织及细胞的收集、运输及保存， CTK40-X1适用于淋巴、血样或骨髓等组织样品的收集、运输及保存。样品体积zui大为8 L。
试剂盒内含：
（1）8L无菌收集盒或管（用于血样等液体样品）
（2）无菌转移液500ml*2瓶: CTK40-X为ZTM:A2™ (Cat. #CTK20-M，含克林霉素，庆大霉素及两性霉素) ； CTK40-X1为ZTM:A1™ (Cat. #CTK20-M1，含克林霉素，庆大霉素)
（3）无菌塑料组织转运袋
（4）各式标签及冰袋
订货信息:
|
货号 |
品名 |
CAS |
价格￥ |
规格 |
品牌 |
|
CTK40-XL |
COLLECTION KITS , X-Large, |
6460 |
INCELL Corporation |
上海金畔生物科技有限公司
杨建辉
上海市浦东绣川路561号1102室
021-50837765 传真 : WWW.78BIO.COM 产品查询 CX.78BIO.COM
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Large Cell and Tissue Collection Kits
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上海金畔生物科技有限公司incell代理，竭诚为您提供incell代理产品，产品*，货到付款，实验无忧！
Large Cell and Tissue Collection Kits
CTK30-L适用于常规组织及细胞的收集、运输及保存，CTK30-L1适用于淋巴、血样或骨髓等组织样品的收集、运输及保存。样品体积zui大为4.5 L。
试剂盒内含：
（1）4.5L无菌收集盒或管（用于血样等液体样品）
（2）无菌转移液500ml*1瓶: Cat. #CTK30-L为ZTM:A2™ (Cat. #CTK20-M，含克林霉素，庆大霉素及两性霉素) ； Cat. #CTK30-L1为ZTM:A1™ (Cat. #CTK20-M1，含克林霉素，庆大霉素)
（3）无菌塑料组织转运袋
（4）各式标签及冰袋
订货信息:
|
货号 |
品名 |
CAS |
价格￥ |
规格 |
品牌 |
|
CTK30-L |
COLLECTION KITS , Large, |
5780 |
INCELL Corporation |
上海金畔生物科技有限公司
杨建辉
上海市浦东绣川路561号1102室
021-50837765 传真 : WWW.78BIO.COM 产品查询 CX.78BIO.COM
我们公司zui大优势是强大的采购，
1：基本什么都能进口，血清，抗体，耗材，还有部分限制进口的，
2： 货品全，现经营过700多个品牌，基本所有生物试剂耗材都可以进口，特别是冷偏的产品那就更有优势，
3：提供加急服务，一般1-2周到货，超过时限加急费全免
4：价格公道，绝大部分价格有优势，当然不能保证100%产品都有价格优势，
5：良好的信誉，大部分客户我们提供货到付款服务，客户包括清华，北大 交大 复旦，中山等100多所大学，ROCHE，阿斯利康，国药 ，fisher等500多家公司
6：我们还是santa,Advanced Biotechnologies Inc;Athens Research & Technology, bangs, BBInternational, crystalchem, dianova, FD Neurotechnologies, Inc. FormuMax Scientific, Inc; Genebridege; Glycotope Biotechnology GmbH; iduron; Innovative Research of America； Ludger ； neuroprobe; omicronbio; Polysciences; prospecbi; QA-BIO；quickzyme；RESEARCH DIETS, INC；sterlitech；sysy；TriLink BioTechnologies, Inc；worthington-biochem;zyagen;……几十家国外公司代理
7：我们还从事invitrogen，qiagen; abcam ;sigma;neb; roche;merck; rnd; BD; GE; pierce; BioLegend….等*批发