T750872	Component	600U	3000 U	Storage
T750872A	T4 RNA Ligase 1	20 μL	100 μL	-20°C. Avoid freeze/thaw cycle
T750872B	10×T4 RNA Ligase 1 Buffer	100 μL	500 μL	-20°C. Avoid freeze/thaw cycle
T750872C	50% PEG 8000	100 μL	500 μL	-20°C. Avoid freeze/thaw cycle
T750872D	ATP	10 μL	50 μL	-20°C. Avoid freeze/thaw cycle

酶活定义
37℃ 条件下，30 min 内将 1 nmol 的 5’-[32P] rA16 转化成磷酸盐不溶物所需要的酶量定义为 1 个单位（U）。

产品使用步骤

1.RNA 的环化反应

(1)按照下表所示配制混合溶液:

Component	Volume
ssRNA	200 ng-1 μg
10×T4 RNA Ligase 1 Buffer	2 μl
ATP	20-50 μM
50% PEG 8000	10% PEG 8000
Murine RNase Inhibitor	0.5-1 μl
T4 RNA Ligase 1	1 μl
Nuclease-free Water	To 20 μl

(2)25 °C 孵育2h或16°C孵育16h。
(3)65 °C 15 min 或煮沸2min，终止反应。
2.RNA 或 DNA 寡核苷酸和 ssRNA 的连接反应
(1)按照下表所示配制混合溶液:

Component	Volume
ssRNA	1-20 pmol
RNA 或 DNA寡核苷酸	5-40 pmol
10×T4 RNA Ligase 1 Buffer	2 μl
ATP	2 μl
50% PEG 8000	15% PEG 8000
Murine RNase Inhibitor	0.5-1 μl
T4 RNA Ligase 1	1 μl
Nuclease-free Water	To 20 μl

(2)25°C孵育2h或16°C孵育16h。
(3)65°C15 min 或煮沸2min，终止反应。
注意事项
本产品仅作科学研究使用，不得用于其它用途。

Storage Buffer： 10mM Tris-HCl, 50mM KCl, 0.1mM EDTA, 1mM DTT, 50% Glycerol, pH 7.5

T750872	Component	600U	3000 U	Storage
T750872A	T4 RNA Ligase 1	20 μL	100 μL	-20°C. Avoid freeze/thaw cycle
T750872B	10×T4 RNA Ligase 1 Buffer	100 μL	500 μL	-20°C. Avoid freeze/thaw cycle
T750872C	50% PEG 8000	100 μL	500 μL	-20°C. Avoid freeze/thaw cycle
T750872D	ATP	10 μL	50 μL	-20°C. Avoid freeze/thaw cycle

T4 RNA Ligase 1 is an ATP-dependent single-stranded nucleic acid molecule (DNA or RNA) ligase that catalyzes the 5' phosphate group of the donor and the 3' hydroxyl group of the ligand to form 3'→5' phosphodiester bonds and accompanies ATP hydrolysis to AMP and PPi. The substrates of this enzyme are ssRNA, ssDNA and dinucleoside pyrophosphate.

Enzyme Activity Definition
At 37℃, the amount of enzyme required to convert 1 nmol of 5 '-[32P] rA16 into phosphate insoluble matter within 30 min is defined as 1 unit (U).
Protocol

1. Cyclization of RNA
(1) Prepare the mixed solution as shown in the following table:

Component	Volume
ssRNA	200 ng-1 μg
10×T4 RNA Ligase 1 Buffer	2 μl
ATP	20-50 μM
50% PEG 8000	10% PEG 8000
Murine RNase Inhibitor	0.5-1 μl
T4 RNA Ligase 1	1 μl
Nuclease-free Water	To 20 μl

(2) Incubation at 25 °C for 2h or 16°C for 16h.
(3) The reaction was terminated by boiling at 65 °C for 15 min or 2min.
2. Linking reaction between RNA or DNA oligonucleotides and ssRNA
(1) Prepare the mixed solution as shown in the following table:

Component	Volume
ssRNA	1-20 pmol
RNA or DNA oligonucleotides	5-40 pmol
10×T4 RNA Ligase 1 Buffer	2 μl
ATP	2 μl
50% PEG 8000	15% PEG 8000
Murine RNase Inhibitor	0.5-1 μl
T4 RNA Ligase 1	1 μl
Nuclease-free Water	To 20 μl

(2) Incubation at 25°C for 2h or 16°C for 16h.
(3) The reaction was terminated at 65°C15 min or by boiling for 2min.
Cautions
This product is for scientific research purposes only and shall not be used for other purposes.

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找到2个结果

批号(Lot Number)	证书类型	货号
ZJ25F0319678	分析证书	T750872
ZJ25F0319677	分析证书	T750872

技术文档和文章

溶液计算器

摩尔浓度计算器

计算溶液所需的质量、体积或浓度。

质量

浓度

体积

分子量

g/mol

稀释计算器

计算配制储备溶液所需的稀释度。

浓度1（C1）

体积 1 (V1)

浓度 2 (C2)

体积 2 (V2)

复溶计算器

复溶计算器允许您快速计算试剂的体积，以复溶小瓶。只需输入试剂的质量和目标浓度，计算器将确定其余部分。

体积（添加到小瓶中）

质量（小瓶）

所需的复溶浓度

T4 RNA Ligase 1

库存信息