Product FAQ

Do I have to use your BLOCK-iT TOPO Transcription Kit to generate the long dsRNA as a template to work with BLOCK-iT Dicer?

Answer

While the BLOCK-iT TOPO Transcription kit is ideal for generating high yields of quality dsRNA for use as a Dicer substrate, long dsRNA from other sources will work as well.

Answer Id: E4419

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Limited Use Label License

BLOCK-iT™ RNAi TOPO® Transcription Kit

Limited Use Label License
172
236
Catalog #
  • K350001(Discontinued)

Product FAQ

What reverse transcription kit should I use for my gene expression experiment?

Answer

Please review this selection table (https://www.thermofisher.com/us/en/home/life-science/pcr/reverse-transcription/cdna-synthesis-kits/rt-real-time-pcr.html) to help choose the best reverse transcription kit for your needs.

Answer Id: E7475

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Manual / Product Insert

User Guide: TranscriptAid T7 High Yield Transcription Kit

Version: Jan. 2015
Catalog #

Product FAQ

Can cDNA be generated using the High Capacity cDNA Reverse Transcription kit (SKU 4322171) for quantitative qPCR applications?

Answer

Yes, the single-stranded cDNA generated via the High-Capacity cDNA Reverse Transcription can be used in real-time PCR applications. In addition, we also recommend Superscript VILO cDNA Synthesis Kit, High Capacity RNA to cDNA kit, and/or High Capacity RNA to cDNA Master Mix for total RNA.

Answer Id: E2149

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Product FAQ

How much RNA do I need for the High-Capacity cDNA Reverse Transcription Kit?

Answer

With the High Capacity Reverse Transcription Kit, you can use from 20 ng up to 2 µg of total RNA in a 20 µL reaction.

Answer Id: E7476

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Product FAQ

I am using the MEGAscript SP6 Transcription Kit for in vitro transcription. I ran my reaction product on a denaturing gel and got a smear. Can you offer some advice?

Answer

If the RNA appears degraded (e.g., smeared), remove residual RNase from the DNA template preparation before in vitro transcription. Do this by digesting the DNA prep with proteinase K (100-200 µg/mL) in the presence of 0.5% SDS for 30 min at 50°C, and follow this with phenol/chloroform extraction. The RNase Inhibitor that is present in the transcription reaction can only inactivate trace RNase contamination. Large amounts of RNase contamination will compromise the size and amount of transcription products.

Answer Id: E9776

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Manual / Product Insert

Protocol: High-Capacity cDNA Reverse Transcription Kits For 200 and 1000 Reactions (English )

Version: 20 Jul 2010
Catalog #

Product FAQ

The DTT in my reverse transcription kit has precipitated—can I still use it?

Answer

No, the DTT will need to be replaced.

Answer Id: E7311

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Product FAQ

Is it possible to generate a full-length cDNA from mRNA attached to Dynabeads magnetic beads, and what reverse transcription kit do you recommend?

Answer

It is possible to generate full-length cDNA from mRNA attached to Dynabeads magnetic beads. We recommend a thermostable reverse transcription kit, so that difficult regions with GC-rich secondary structures are accommodated. However, it is not possible to start the reaction by heating the mRNA on the beads because that will elute the mRNA (A:T base pairs are the least thermostable).

We have used ThermoScript reverse transcriptase, inhouse, with Oligo(dT)25 on the beads as primers. The cDNA synthesis was performed according to the manufacturer's instructions. When using a thermostable reverse transcriptase and the Oligo(dT)25 as primer for first-strand cDNA synthesis, an initial step of incubation at 50 degrees C for 5 min is necessary before proceeding at the recommended elevated temperature. This is to start the cDNA synthesis beyond the A:T hybridization point so that the mRNA doesn't fall off the beads. The resulting cDNA is covalently attached to the bead surface, and the beads with the attached cDNA can be used as template in multiple hybridization reactions.

Answer Id: E6078

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Product FAQ

What is the difference between the High-Capacity RNA-to-cDNA Kit, the High-Capacity cDNA Reverse Transcription Kit, and the SuperScript VILO Master Mix?

Answer

The main differences are in the enzyme and type of reverse transcription primer used. The High-Capacity kits use MultiScribe Reverse Transcriptase, while the SuperScript VILO Master Mix uses SuperScript III Reverse Transcriptase. The High-Capacity RNA-to-cDNA Kit uses a blend of random primers and oligo(dT), while the High-Capacity cDNA Reverse Transcription Kit and the SuperScript VILO Master Mix use only random primers.

Answer Id: E7474

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Product FAQ

What is the advantage of using TranscriptAid T7 High Yield Transcription Kit over Thermo Scientific T7 RNA Polymerase with a standard transcription buffer?

Answer

TranscriptAid T7 High Yield Transcription Kit is designed to produce both long and short transcripts for applications that require large yields of RNA (milligram amounts). The kit can achieve 10 times greater amount than from conventional in vitro transcription reactions with T7 RNA polymerase due to its unique proprietary enzyme and buffer formulations. Please note that TranscriptAid T7 High Yield Transcription Kit is not recommended for generation of radioactively labelled RNA. Due to large quantities of RNA synthesized with the kit, generation of high specific activity radiolabeled probes would require prohibitively large amounts of radiolabeled nucleotide.

Answer Id: E8839

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Manual / Product Insert

BLOCK-iT RNAi TOPO Transcription Kit

Version: 4 January 2012
Catalog #
  • K350001(Discontinued)
  • K365001(Discontinued)

Manual / Product Insert

PI: High-Capacity cDNA Reverse Transcription Kits

Version: 4375222 Rev C (21Mar2016)