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分子生物学
核酸标记和检测
T4 DNA Ligase for NGS

T4 DNA Ligase for NGS

COA

有货

库存信息

关闭

库存信息

关闭

货号 (SKU)	包装规格	是否现货	价格	数量
T665501-1500U	1500U	期货
T665501-7500U	7500U	期货

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基本描述

别名

用于NGS的T4 DNA连接酶

英文名称

T4 DNA Ligase for NGS

储存温度

-20°C储存,避免反复冻融

运输条件

超低温冰袋运输

产品介绍

T4 DNA Ligase是从表达T4 DNA Ligase基因的大肠杆菌经诱导表达后分离纯化而来的，催化相邻DNA链的5’磷酸基团和3’羟基基团以磷酸二酯键结合反应。该酶可催化平末端或粘性末端DNA的连接，修复双链DNA、RNA、DNA/RNA杂交中的单链中的单链切口，但是对于单链核苷酸，没有活性。

T665501	Component	1500 U	7500 U	Storage
T665501A	T4 DNA Ligase, 15 U/μL	100 μL	500 μL	-20°C. Avoid freeze/thaw cycle.
T665501B	4×T4 DNA Ligase Buffer	600 μL	2×1.5 mL	-20°C. Avoid freeze/thaw cycle.

活性定义:

1U是指在ATP-PPi交换反应中，37°C、20分钟内将1nmol [32PPi]转换为Norit可吸收形式所需的酶量，相当于约 200个粘性末端连接单位。

应用范围:

主要用于NGS中文库构建过程中的Adaptor连接。

使用方法:

建议使用Adaptor进行连接，也可选择使用NEB、Illumina公司的Adaptor，具体连接方法可参考各公司的产品使用说明书。以下为使用Adaptor进行连接的操作步骤：

1．向已完成DNA末端修复的反应液中直接加入以下试剂：

此时管中溶液总体积为100 μL。

注意：若起始样本量少于100 ng，请将Adaptor用去离子水稀释10倍至1.5 μM后使用。

2．用枪上述试剂吹吸混匀，短暂离心，使溶液收集到管底。

3．23℃温浴20分钟。

注意：若此操作使用PCR仪，请将热盖关闭。

4．继续进行后续步骤，如DNA片段的选择性回收或DNA片段的纯化。

T4 DNA Ligase is isolated and purified from Escherichia coli expressing the T4 DNA Ligase gene after induction and expression. It catalyzes the binding reaction of adjacent DNA strands with 5 'phosphate groups and 3' hydroxyl groups via phosphodiester bonds. This enzyme can catalyze the connection of flat or sticky end DNA, repairing single stranded incisions in double stranded DNA, RNA, and DNA/RNA hybridization, but has no activity for single stranded nucleotides.

T665501	Component	1500 U	7500 U	Storage
T665501A	T4 DNA Ligase, 15 U/μL	100 μL	500 μL	-20°C. Avoid freeze/thaw cycle.
T665501B	4×T4 DNA Ligase Buffer	600 μL	2×1.5 mL	-20°C. Avoid freeze/thaw cycle.

Activity definition:

1U refers to the amount of enzyme required to convert 1nmol [32PPi] into Norit absorbable form within 20 minutes at 37 ° C in the ATP-PPi exchange reaction, equivalent to approximately 200 viscous terminal junction units.

Application scope:

Mainly used for adapter connection during the construction process of NGS Chinese library.

Usage:

It is recommended to use Kangwei Adaptor for connection, or choose to use Adaptors from NEB or Illumina companies. For specific connection methods, please refer to the product manuals of each company. The following are the steps to connect using Kangwei Adaptor:
1. Directly add the following reagents to the reaction solution that has completed DNA end repair:

Reagents Name	volume
4×T4 DNA ligase Buffer	25 μL
T4 DNA ligase ,15 U/μL	5 μL
Adaptor	5 μL
ddH2 O	Supplemented to 50 μ L

At this point, the total volume of solution in the tube is 100 μ L.
Note: If the initial sample size is less than 100 ng, please dilute the adapter 10 times with deionized water to 1.5 μ Use after M.
2. Use a gun to blow and aspirate the above reagents, mix well, and centrifuge briefly to collect the solution to the bottom of the tube.
Take a warm bath at

3.23 ℃ for 20 minutes.
Attention: If using a PCR instrument for this operation, please close the heat cap.
4. Continue with subsequent steps, such as selective recovery of DNA fragments or purification of DNA fragments.