RNA-related products, FAQ

PowerCut™ Dicer

Q 01: What is the origin of PowerCut Dicer?
PowerCut Dicer is a recombinant Dicer originating from Giardia intestinalis. The protein only contains the domains that are necessary for the in vitro cleaving activity for a Dicer protein.

Q 02: What size of fragments does PowerCut Dicer produce?
PowerCut Dicer cleaves the dsRNA substrate every 25-27 nt, generating siRNA molecules 23-25 bp long with 2 nt overhangs in the 3’ end.

Q 03: What size of dsRNA can I use as a substrate with PowerCut Dicer?
There are no known size limitations for the PowerCut Dicer substrate. We have successfully cleaved substrates up to 6 kb in length.

Q 04: Can I cleave more dsRNA substrate in a reaction if I extend the reaction time?
We do not recommend extending the reaction time or using much more substrate than 1 µg/1U PowerCut Dicer. Adding more than the recommended amount of substrate will probably decrease the yield of the PowerCut Dicer reaction.

Q 05: Why does the PowerCut Dicer not cleave my substrate?
Make sure you have followed the reaction condition guidelines if you have scaled the raction up or down. Also, make sure that there are no inactivating components transferred to the reaction with the dsRNA substrate. Inhibiting components like EDTA and LiCl can be transferred to the reaction with dsRNA if they are carried over from purification or precipitation of the dsRNA.

Q 06: Do I need to purify my synthesized dsRNA before using it as a substrate in the PowerCut Dicer reaction?
We strongly recommend purification of the dsRNA synthesis reaction before using the dsRNA as a substrate, as synthesis reactions may contain components that can interfere with PowerCut Dicer activity. Purifying the dsRNA will also make spectrophotometric measurements of the concentration accurate, since unincorporated ribonucleotides, DNA template and unwanted RNA products are separated from the produced dsRNA.

Q 07: What kind of purification protocol can I use to purify the dsRNA substrate before using it in PowerCut Dicer reaction?
We recommend using LiCl precipitation to purify the dsRNA substrate. A suitable protocol can be found in the Technical appendix for RNA-related products.

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