Hotstart HiTaq One-Step RT-PCR Mix(Heat-labile UDG)

品牌：Jinpan

Hotstart HiTaq One-Step RT-PCR Mix(Heat-labile UDG)

MSDS

质检证书(CoA)

相似产品

货号 (SKU)	包装规格	是否现货	价格	数量
H292677-50μl	50μL	期货

产品名称	Hotstart HiTaq One-Step RT-PCR Mix(Heat-labile UDG)
英文名称	Hotstart HiTaq One-Step RT-PCR Mix(Heat-labile UDG)
运输条件	超低温冰袋运输

产品简介

One-Step  RT-PCR  Mix(Heat-labile  UDG)以其卓越的快速性及定量性被广泛应用，使用该产品进行 Real Time RT-PCR 反应可在同一反应管内连续进行，操作简单，并能有效防止污染。本制品中使用了最适合于 Real Time RT-PCR 的超级反转录酶（Super MMLV Reverse Transcriptase）和化学修饰热启动酶（Hotstart HiTaq DNA Polymerase），具有高扩增效率和高扩增特异性。本试剂盒中引入了 dUTP/UDG 防污染系统，Heat-labile UDG 在室温下即可将含U 的污染物迅速降解，55℃逆转录时，Heat-labile UDG迅速失活，因此不会影响 RT-PCR 的效率和灵敏度。

dUTP/UDG 防污染系统是一种非常有效的控制 PCR 扩增产物污染的手段。含有比例优化的dUTP/dNTP  Mix，并加入了来源于嗜冷海洋细菌的  Heat-labile  Uracil-DNA  Glycosylase。Heat-labile Uracil-DNA Glycosylase 在室温下具有很高的活性，在反应体系混合过程中即可充分降解含U 的双链DNA污染物。当反应体系升温至 55℃  时，Heat-labile Uracil-DNA Glycosylase 迅速彻底失活，维持 cDNA 的完整性，保证检测灵敏度不受影响。

产品特点

快速方便的单管反应；

适合各种RNA 模板，无需优化反应条件；

独特的酶组合，确保反应的高特异性和高灵敏度；

经优化的缓冲体系确保了高效的逆转录和扩增过程。

产品内容

Hotstart HiTaq DNA Polymerase ( 5U/μl )

Super MMLV Reverse Transcriptase ( 200 U /μl )

RNasin ( 40U/μl )

5×HS HiTaq One Step RT-PCR Buffer(Mg2+ Plus)

Solution I ( 10× )

dN(U)TP Mix ( 20mM，A:C:G:T:U=1:1:1:1:1 )

Heat-labile Uracil-DNA Glycosylase(1U/μl)

使用方法

1. 体系配制（50μl 体系）

组分	体积	终浓度
Hotstart HiTaq DNA Polymerase	0.375μl	—
Super MMLV Reverse Transcriptase	0.2μl	—
RNasin	0.25μl	—
5×HS HiTaq One Step RT-PCR	5μl	1×
Solution I ( 10× )	2.5μl	1×
dN(U)TP Mix ( 20mM )	0.25μl	0.2mM
Heat-labile Uracil-DNA Glycosylase	0.25μl	—
Primer-probe Mix	1.5μl ※	—
模板	2.5μl ※	—
超纯水	Up to 25μl	—
总体积	25μl	—

※ 注：引物、探针以及模板的用量可自行调整，体系不够请用超纯水补齐。

2.反应条件

体系配制完成后，室温放置 15min 后，上机进行检测：

步骤	温度	时间	循环数
1	50℃	15min	1
2	95℃	10min	1
3	94℃	15s	40※
4	55℃	40s（收集荧光）

※注：反应条件根据引物探针和模板情况可自行设置。

Product Introduction

One-Step RT-PCR Mix (Heat-labile UDG) is widely used due to its excellent rapidity and quantification. Real Time RT-PCR reactions can be carried out continuously in the same reaction tube with this product, which is simple and effective. Prevent pollution. This product uses Super MMLV Reverse Transcriptase (Super MMLV Reverse Transcriptase) and chemically modified Hotstart HiTaq DNA Polymerase, which are most suitable for Real Time RT-PCR, with high amplification efficiency and high amplification specificity. The dUTP/UDG anti-pollution system is introduced in this kit. Heat-labile UDG can rapidly degrade U-containing contaminants at room temperature. When reverse transcription at 55°C, Heat-labile UDG is rapidly inactivated, so it will not affect RT -The efficiency and sensitivity of PCR.

The dUTP/UDG anti-pollution system is a very effective means to control the contamination of PCR products. Contains optimized dUTP/dNTP Mix and added Heat-labile Uracil-DNA Glycosylase derived from psychrophilic marine bacteria. Heat-labile Uracil-DNA Glycosylase has high activity at room temperature, and it can fully degrade U-containing double-stranded DNA contaminants during the mixing process of the reaction system. When the reaction system is heated to 55°C, Heat-labile Uracil-DNA Glycosylase is quickly and completely inactivated, maintaining the integrity of the cDNA and ensuring that the detection sensitivity is not affected.

Features

Quick and convenient single tube reaction;

Suitable for various RNA templates, no need to optimize reaction conditions;

Unique enzyme combination to ensure high specificity and high sensitivity of the reaction;

The optimized buffer system ensures an efficient reverse transcription and amplification process.

Product content

Hotstart HiTaq DNA Polymerase ( 5U/μl )

Super MMLV Reverse Transcriptase ( 200 U /μl )

RNasin ( 40U/μl )

5×HS HiTaq One Step RT-PCR Buffer(Mg2+ Plus)

Solution I ( 10× )

dN(U)TP Mix ( 20mM，A:C:G:T:U=1:1:1:1:1 )

Heat-labile Uracil-DNA Glycosylase(1U/μl)

Instructions

1. System preparation (50μl system)

Reagent	Volume	Final concentration
Hotstart HiTaq DNA Polymerase	0.375μl	—
Super MMLV Reverse Transcriptase	0.2μl	—
RNasin	0.25μl	—
5×HS HiTaq One Step RT-PCR	5μl	1×
Solution I ( 10× )	2.5μl	1×
dN(U)TP Mix ( 20mM )	0.25μl	0.2mM
Heat-labile Uracil-DNA Glycosylase	0.25μl	—
Primer-probe Mix	1.5μl ※	—
Template	2.5μl ※	—
Ultra-pure water	Up to 25μl	—
Total capacity	25μl	—

※ Note: The amount of primers, probes and templates can be adjusted by yourself. If the system is not enough, please use ultrapure water to make up.

2. Reaction conditions

After the preparation of the system is completed, leave it at room temperature for 15 minutes, and then test it on the machine:

Step	Temperature	Time	Number of cycles
1	50℃	15min	1
2	95℃	10min	1
3	94℃	15s	40※
4	55℃	40s(Collect fluorescence)

※Note: The reaction conditions can be set according to the primer probe and template.

储存温度	-20°C储存
品牌	Jinpan