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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