货号:L6030-HC-L
品名:T4 DNA Ligase (Rapid) (T4 DNA连接酶(快速))
产品描述:
T4 DNA连接酶催化双链DNA或RNA的末端5'磷酸和3'羟基之间形成磷酸二酯键。该酶有效连接钝端和粘端,并修复双链DNA,RNA或DNA / RNA杂合体中的单链缺口(1)。
规格:
|
600,000 U/ml
|
浓度:
|
240,000 U
|
保存:
|
-25°C至-15°C
|
纯度(SDS-PAGE)
|
> 99%
|
活性:
|
300,000 U/mg
|
SS核酸外切酶
|
6,000 U< 1.0 % Released
|
DS核酸外切酶
|
6,000 U< 1.0 % Released
|
DS内切核酸酶
|
6,000 U=No Conversion
|
大肠杆菌 DNA污染
|
6,000 U <10 copies
|
蛋白质来源
携带克隆的T4 DNA连接酶基因的重组大肠杆菌菌株。
提供:
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% glycerol
pH 7.5 @ 25°C
随附:
B1010 (2X Rapid Ligation Buffer)
B6030 (10X T4 DNA Ligase Buffer)
2X Rapid Ligation Buffer (B1010):
132mM Tris-HCI
20 mM MgCl2
2 mM DTT
2 mM ATP
15% PEG 6000
pH 7.6 @ 25°C
10X T4 DNA Ligase Buffer (B6030):
500mM Tris-HCI
100 mM MgCl2
50 mM DTT
10 mM ATP
pH 7.6 @ 25°C
单位定义
1单位定义为在23℃孵育30分钟后,在50μl1X DNA连接酶缓冲液中加入100ng具有粘性末端的DNA片段的50%所需的DNA连接酶的量。
使用说明:
1、反应设置
Amount
|
Description
|
Final Concentration
|
10 uL
|
2X Rapid Ligation Buffer
|
1X
|
X uL
|
Vector
|
1-10 ng/uL
|
X uL
|
Insert
|
1-10 ng/uL
|
1 uL
|
T4 DNA Ligase (600 U/ uL)
|
30 U/uL
|
X uL
|
Type I Water
|
N/A
|
20 uL
|
Total Volume
|
|
2、过程操作
1)将所有上述组分加入干净的反应容器中,通过移液充分混合。
2)25°C孵育10分钟。
3)使用PCR清洁柱立即纯化DNA并洗脱约50μL。
4)– 或 – 在TE或水中立即稀释(至少1:10,但足以使0.1-10ng连接产物转化)
5)将0.1-10ng连接产物转化为与载体相容的化学或电子感受态细胞系
3、其他
在1X T4 DNA连接酶反应缓冲液中用Lambda-Hind III DNA片段底物测量,一个酶酶T4 DNA连接酶粘性末端单位相当于约3个粘性末端单位。
一个Weiss单位大约相当于22个Enzymatics粘性末端单位。
T4 DNA连接酶是ATP依赖性的。建议在-20°C储存一年后丢弃反应缓冲液,并用新鲜缓冲液代替以确保*性能。当靶向插入片段:载体比率在2和6之间时,单插入连接是*的。高于6:1的比率将促进插入多个片段,而降至2:1以下将降低连接效率。
对于有问题的连接或DNA浓度未知的情况,可能需要改变比率并进行多次连接。
高浓度PEG的存在将显着降低电感受态细胞的转化效率。为了zui大限度地转化为电感受态细胞,建议采用以下方法:
*: 连接后,使用DNA纯化离心柱纯化产物并在50μLTE中洗脱。DNA现在已准备好转化。要转化的DNA的zui终数量应该在0.1-10ng范围内。
更好: 在ddH2O或TE中稀释连接产物以降低PEG浓度。要转化的DNA的zui终数量应该在0.1-10ng范围内。
Enzymatics的高浓度T4 DNA连接酶与2X快速连接缓冲液结合使用,极大地刺激了平端连接的速度和效率,因此不建议长时间孵育(> 10分钟),并可能大大降低连接产物的转化效率。为了使正确的插入/载体组合的转化效率zui大化,推荐使用以下方案。
酶学10X T4 DNA连接酶缓冲液不含PEG,并且与标准连接方案相兼容,该方案不规定使用快速/快速/快速格式缓冲液。
部分产品信息:
货号
|
品名
|
规格
|
浓度
|
品牌
|
N2070-10-L
|
ATP Solution
|
50 µmol
|
10 mM
|
Enzymatics
|
N2050-10-L
|
10 mM dNTP Solution Mix
|
100 µmol
|
10 mM
|
Enzymatics
|
N2010L
|
100 mM 75µmol dNTP Solution Set
|
75 µmol
|
100 mM
|
Enzymatics
|
N2010-A-L
|
100 mM dATP Solution
|
75 µmol
|
100 mM
|
Enzymatics
|
N2050L
|
100µmol dNTP Solution Mix
|
100 µmol
|
25 mM
|
Enzymatics
|
Y9180L
|
10x Uracil Cleavage System
|
750 Reactions
|
N/A
|
Enzymatics
|
P7670
|
2x HiFi PCR Master Mix
|
1 rxn
|
2x
|
Enzymatics
|
Y9420L
|
5x ER/A-Tailing Enzyme Mix
|
24 reactions
|
N/A
|
Enzymatics
|
Y9380L
|
E. coli Pyrophosphatase
|
50 U
|
100 U/mL
|
Enzymatics
|
Y9030L
|
E. coli Single-Stranded DNA Binding Protein
|
1.5 mg
|
2.75 mg/ml
|
Enzymatics
|
Y9140-HC-L
|
End-Repair Mix (High Concentration)
|
75 Reactions
|
1 Rxn/µL
|
Enzymatics
|
Y9140-LC-L
|
End-Repair Mix (Low Concentration)
|
200 Reactions
|
1 Rxn/µL
|
Enzymatics
|
Y9080L
|
Endonuclease VIII
|
10,000 U
|
10,000 U/ml
|
Enzymatics
|
P7600L
|
EnzScript™ (MMLV Reverse Transcriptase RNase H-)
|
10,000 U
|
200,000 U/mL
|
Enzymatics
|
X8010L
|
Exonuclease I
|
30,000 U
|
20,000 U/ml
|
Enzymatics
|
X8020L
|
Exonuclease III
|
50,000 U
|
100,000 U/ml
|
Enzymatics
|
P7010-HC-L
|
Klenow (3′→ 5′ exo-) (High Concentration)
|
10,000 U
|
50,000 U/ml
|
Enzymatics
|
P7010-LC-L
|
Klenow (3′→ 5′ exo-) (Low Concentration)
|
10,000 U
|
5,000 U/ml
|
Enzymatics
|
P7060L
|
Klenow Fragment
|
2,500 U
|
5,000 U/ml
|
Enzymatics
|
X8030L
|
Lambda Exonuclease
|
10,000 U
|
5,000 U/ml
|
Enzymatics
|
P7040L
|
M-MuLV Reverse Transcriptase
|
100,000 U
|
200,000 U/ml
|
Enzymatics
|
P7090L
|
Mako DNA Polymerase (3′→ 5′ exo-)
|
3,000 U
|
30,000 U/ml
|
Enzymatics
|
P7140-HC-L
|
Manta 1.0 DNA Polymerase (High Concentration)
|
100,000 U
|
400,000 U/ml
|
Enzymatics
|
P7140-LC-L
|
Manta 1.0 DNA Polymerase (Low Concentration)
|
100,000 U
|
40,000 U/ml
|
Enzymatics
|
P7610L
|
NGS Library Prep: 2X VeraSeq™ PCR Mix – 250 Reactions
|
250 Reactions
|
2X
|
Enzymatics
|
P7590L
|
Phoenix™ Hot Start Taq DNA Polymerase
|
500 U
|
5,000 U/ml
|
Enzymatics
|
P7460L
|
Poly(A) Polymerase
|
1,000 U
|
5,000 U/mL
|
Enzymatics
|
Y9260L
|
RecA
|
1.5 mg
|
2.0 mg/mL
|
Enzymatics
|
Y9220L
|
RNAse H
|
5,000 U
|
5,000 U/ml
|
Enzymatics
|
Y9240L
|
RNAse Inhibitor
|
20,000 U
|
40,000 U/ml
|
Enzymatics
|
L6010L
|
T3 DNA Ligase
|
900,000 U
|
3,000,000 U/ml
|
Enzymatics
|
L6030-LC-L
|
T4 DNA Ligase
|
150,000 U
|
120,000 U/ml
|
Enzymatics
|
L6030-HC-L
|
T4 DNA Ligase (Rapid)
|
240,000 U
|
600,000 U/ml
|
Enzymatics
|
P7080L
|
T4 DNA Polymerase
|
2,000 U
|
3,000 U/ml
|
Enzymatics
|
Y9130L
|
T4 Gene 32 Protein
|
1.0 mg
|
10 mg/mL
|
Enzymatics
|
Y9040L
|
T4 Polynucleotide Kinase
|
10,000 U
|
10,000 U/ml
|
Enzymatics
|
L6050L
|
T4 RNA Ligase 1
|
10,000 U
|
20,000 U/ml
|
Enzymatics
|
L6080L
|
T4 RNA Ligase 2
|
4,500 U
|
30,000 U/ml
|
Enzymatics
|
L6070L
|
T4 RNA Ligase 2 Truncated
|
500 U
|
5,000 U/ml
|
Enzymatics
|
L6020L
|
T7 DNA Ligase
|
900,000 U
|
3,000,000 U/ml
|
Enzymatics
|
P7260L
|
T7 DNA Polymerase
|
3,500 U
|
10,000 U/ml
|
Enzymatics
|
P7180L
|
T7 RNA Polymerase
|
50,000 U
|
50,000 U/ml
|
Enzymatics
|
L6060L
|
Taq DNA Ligase
|
20,000 U
|
40,000 U/ml
|
Enzymatics
|
P7110L
|
Taq DSC 2.0 DNA Polymerase
|
1,250 U
|
5,000 U/ml
|
Enzymatics
|
P7250L
|
Taq-B DNA Polymerase
|
10,000 U
|
5,000 U/ml
|
Enzymatics
|
P7620L
|
TaqIT
|
5,000 U
|
50,000 U/mL
|
Enzymatics
|
P7070L
|
Terminal deoxynucleotidyl Transferase (TdT)
|
6,000 U
|
20,000 U/ml
|
Enzymatics
|
G5020L
|
Thermolabile UDG
|
500 U
|
1,000 U/ml
|
Enzymatics
|
Y9370L
|
Thermostable Pyrophosphatase
|
1,250 U
|
2,000 U/ml
|
Enzymatics
|
G5010L
|
Uracil DNA Glycosylase
|
10,000 U
|
2,000 U/ml
|
Enzymatics
|
P7511L
|
VeraSeq™ 2.0 High-Fidelity DNA Polymerase
|
500 U
|
2,000 U/ml
|
Enzymatics
|
P7520L
|
VeraSeq™ ULtra DNA Polymerase
|
500 U
|
2,000 U/ml
|
Enzymatics
|
L6030-W-L
|
WGS Ligase
|
24 Reactions
|
N/A
|
Enzymatics
|
P7640L
|
ZipScript™ One-Step RT-qPCR Kit
|
1000 Reactions
|
N/A
|
Enzymatics
|
P7020-HC-L
|
φ29 DNA Polymerase (High Concentration)
|
2,000 U
|
100,000 U/ml
|
Enzymatics
|
P7020-LC-L
|
φ29 DNA Polymerase (Low Concentration)
|
2,000 U
|
10,000 U/ml
|
Enzymatics
|
世界*实验材料供应商 Enzymatics上海金畔生物为其中国代理, Enzymatics在一直是行业的*,一直为广大科研客户提供zui为的产品和服务,上海金畔生物一直秉承为中国科研客户带来的产品,的服务, Enzymatics就是为了给广大科研客户带来更加完善的产品和服务,您的满意将是我们zui大的收获
Enzymatics中国代理, Enzymatics上海代理, Enzymatics北京代理,Enzymatics广东代理, Enzymatics江苏代理Enzymatics湖北代理,Enzymatics天津,Enzymatics黑龙江代理,Enzymatics内蒙古代理,Enzymatics吉林代理,Enzymatics福建代理, Enzymatics江苏代理, Enzymatics浙江代理, Enzymatics四川代理,
酶产品在生命科学研究中是*的重要工具,在科学研究中对于其性能、纯度和稳定性的要求日益提升。基因公司*代理的酶产品专家—Enzymatics一贯为客户提供zui高质量和*价值的产品,因此我们竭诚向您推荐 Enzymatics产品。
Enzymatics公司建立于2006年,专门从事开发、制造和分销酶产品,致力于酶产品研发,为市场带来了特殊的、高品质的产品。 Enzymatics公司建立了严格完整的操作体系,通过了ISO13485:2003和9001:2000体系标准认证,并符合FDA 21 CFR.820质量系统法规中对于常规用途试剂的生产规范。同时Enzymatics的生产能力被许多FDA认证的医疗设备制造商以及的ISO认证机构TüV SüD 认可。