SuperScript™ VILO™ cDNA Synthesis Kit, 50 reactions - FAQs

SuperScript III反转录酶和SuperScript VILO试剂盒之中的反转录酶有何区别？

SuperScript VILO cDNA合成试剂盒含有SuperScript III反转录酶和辅助蛋白的混合物，其中后者可以提高反转录反应的效率，进而提高得率。由于存在辅助蛋白，SuperScript VILO中的反转录在42°C条件下活化。

PCR反应要加入多少第一链cDNA反应产物？

尽管实际体积取决于用于第一链合成反应的RNA起始量和靶标基因的丰度，但我们建议使用10%的第一链cDNA反应产物进行PCR反应。

How long can I store the cDNA from my reverse transcription step?

You can store your cDNA at 2-6 degrees C for up to 24 hours. For long-term storage, store the cDNA at -15 to -25 degrees C and add EDTA to a final concentration of 1 mM to prevent degradation.

What is the difference between SuperScript III RT and the RT in the SuperScript VILO kit?

The SuperScript VILO cDNA Synthesis Kit contains a mix of SuperScript III RT and helper proteins which help to increase the efficiency of the reverse transcription reaction and thus improve yield. The RT in the SuperScript

The SuperScript VILO cDNA Synthesis Kit (Cat. No. 11754050) contains a mix of SuperScript III RT and helper proteins which help to increase the efficiency of the reverse transcription reaction and thus improve yield. The RT in the SuperScript VILO kit is active at 42 degrees C due to the helper proteins.

How much of the first-strand cDNA reaction should I load for PCR?

While the volume is dependent on the starting amount of RNA used for the first-strand synthesis and the abundance of the target gene, we'd recommend starting with 10% of the first-strand reaction for your PCR reaction.

In what concentration range of total RNA does the SuperScript VILO cDNA Synthesis Kit (Cat. No. 11754050) work?

What is the RNA input for the Oncomine Focus Assay, Chef-Ready Library kit?

Each reverse transcription reaction requires 10 ng of DNase-treated total RNA (≥1.43 ng/µL).

With the Oncomine Focus Assay, Chef-Ready Library, can I perform reverse transcription using the SuperScript VILO cDNA Synthesis Kit on the Ion Chef System?

Reverse transcription needs to be performed manually in the IonCode 96-well PCR plate, using the SuperScript VILO cDNA Synthesis Kit. SuperScript reverse transcriptase enzyme is not supplied in the kit and must be purchased separately. If the RNA was prepared from FFPE tissue and not previously heat-treated, we recommend heating at 80 degrees C for 10 min, then cooling to room temperature.