Hifair® Ⅲ第1链cDNA合成即用型预混液|Hifair® Ⅲ 1st Strand cDNA Synthesis SuperMix

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Hifair® Ⅲ第1链cDNA合成即用型预混液|Hifair® Ⅲ 1st Strand cDNA Synthesis SuperMix

产品说明书

FAQ

COA

已发表文献

Hifair® Ⅲ 1st Strand cDNA Synthesis SuperMix基于Hifair® Ⅲ Reverse Transcriptase而开发的即用型预混液。与Hifair® Ⅱ Reverse Transcriptase相比，Hifair® Ⅲ Reverse Transcriptase热稳定性大幅度提高，可耐受高达60℃的反应温度，适合具有复杂二级结构的RNA模板的逆转录。同时，该酶增强了与模板的亲和力，非常适合少量模板以及低拷贝基因的逆转录。

Hifair® Ⅲ 1st Strand cDNA Synthesis SuperMix为即用型预混液，包含Hifair® Ⅲ Reverse Transcriptase，RNase inhibitor，dNTP，Random primers/Oligo dT primer mix和优化的缓冲体系，只需再加入模板RNA和RNase-free ddH2O即可进行反应。

产品组分

组分编号

组分名称

产品编号/规格

11137ES10 (10 T)

11137ES60 (100 T)

11137-A

RNase-free H2O

1 mL

2×1 mL

11137-B

4×Hifair® Ⅲ SuperMix

50 μL

500 μL

 

产品应用

适用于 RT-qPCR 实验。

 

运输和保存方法

冰袋运输。-20ºC保存，有效期18个月。

 

第一链 cDNA 合成操作步骤

逆转录反应体系

逆转录程序

组分

使用量

温度

时间

RNase-free H2O

To 20 μL

25℃

5 min

4×Hifair® Ⅲ SuperMix

5 μL

55℃

15 min

Total RNA

10 pg -5 μg

85℃

5 min

or mRNA

10 pg-500 ng

    

【注】：1. 20 μL逆转录反应体系建议Total RNA的投入量不超过1 μg。如果目的基因的表达丰度低，最多投入5 μg Total RNA；

2. 逆转录温度：推荐使用55℃。对于高GC含量模板或者复杂模板，可将逆转录温度提高到60℃。

※ 逆转录产物可立即用于后续qPCR反应，也可-20℃短期保存，若需长期保存，建议分装后，于-80℃保存，避免反复冻融。

 

注意事项

1．可以室温溶解，溶解后宜存放于冰盒内或冰浴上，使用完毕后应立即置于-20℃保存。

2．为了您的安全和健康，请穿实验服并戴一次性手套操作。

3．本产品仅作科研用途！ 

HB220308

 

Q：逆转录试剂 11137ES 中引物是随机引物还是oligo dT，还是二者的混合物？

A：二者的混合物。

Q：能否用来做miRNA/circRNA/lncRNA 的逆转录？

A：有A 尾的 lncRNA 可以逆转录， circRNA miRNA 和无 A 尾的 lncRNA 用 不能进行逆转录。microRNA 需要特殊的茎环引物， 需特别针对的逆转录试剂盒。

Q：11137 可以逆转真菌（或其他物种）RNA 吗？

A：RNA 的物种与逆转录没有很大关系，RNA 质量与逆转录有关。所以，得到 RNA 后，逆转录试剂盒都可以用，没有特别针对的。

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Hifair® Ⅲ 1st Strand cDNA Synthesis SuperMix基于Hifair® Ⅲ Reverse Transcriptase而开发的即用型预混液。与Hifair® Ⅱ Reverse Transcriptase相比，Hifair® Ⅲ Reverse Transcriptase热稳定性大幅度提高，可耐受高达60℃的反应温度，适合具有复杂二级结构的RNA模板的逆转录。同时，该酶增强了与模板的亲和力，非常适合少量模板以及低拷贝基因的逆转录。

Hifair® Ⅲ 1st Strand cDNA Synthesis SuperMix为即用型预混液，包含Hifair® Ⅲ Reverse Transcriptase，RNase inhibitor，dNTP，Random primers/Oligo dT primer mix和优化的缓冲体系，只需再加入模板RNA和RNase-free ddH2O即可进行反应。

产品组分

组分编号

组分名称

产品编号/规格

11137ES10 (10 T)

11137ES60 (100 T)

11137-A

RNase-free H2O

1 mL

2×1 mL

11137-B

4×Hifair® Ⅲ SuperMix

50 μL

500 μL

 

产品应用

适用于 RT-qPCR 实验。

 

运输和保存方法

冰袋运输。-20ºC保存，有效期18个月。

 

第一链 cDNA 合成操作步骤

逆转录反应体系

逆转录程序

组分

使用量

温度

时间

RNase-free H2O

To 20 μL

25℃

5 min

4×Hifair® Ⅲ SuperMix

5 μL

55℃

15 min

Total RNA

10 pg -5 μg

85℃

5 min

or mRNA

10 pg-500 ng

    

【注】：1. 20 μL逆转录反应体系建议Total RNA的投入量不超过1 μg。如果目的基因的表达丰度低，最多投入5 μg Total RNA；

2. 逆转录温度：推荐使用55℃。对于高GC含量模板或者复杂模板，可将逆转录温度提高到60℃。

※ 逆转录产物可立即用于后续qPCR反应，也可-20℃短期保存，若需长期保存，建议分装后，于-80℃保存，避免反复冻融。

 

注意事项

1．可以室温溶解，溶解后宜存放于冰盒内或冰浴上，使用完毕后应立即置于-20℃保存。

2．为了您的安全和健康，请穿实验服并戴一次性手套操作。

3．本产品仅作科研用途！ 

HB220308

 

Q：逆转录试剂 11137ES 中引物是随机引物还是oligo dT，还是二者的混合物？

A：二者的混合物。

Q：能否用来做miRNA/circRNA/lncRNA 的逆转录？

A：有A 尾的 lncRNA 可以逆转录， circRNA miRNA 和无 A 尾的 lncRNA 用 不能进行逆转录。microRNA 需要特殊的茎环引物， 需特别针对的逆转录试剂盒。

Q：11137 可以逆转真菌（或其他物种）RNA 吗？

A：RNA 的物种与逆转录没有很大关系，RNA 质量与逆转录有关。所以，得到 RNA 后，逆转录试剂盒都可以用，没有特别针对的。

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