TURBO DNase™ (2 U/μL), 1000 U - FAQs

View additional product information for TURBO DNase™ (2 U/μL) - FAQs (AM2239, AM2238)

9 product FAQs found

How do I remove DNase from an RNA sample?

DNase can be removed by the following methods: acid phenol:choloform extraction, lithium chloride precipitation, EDTA, and heat inactivation, or using our MegaClear Transcription Clean-Up Kit (Cat. No. AM1908).

Alternatively, our Invitrogen TURBO DNA-free Kit (Cat. No. AM1907) is supplied with a DNase inactivation reagent that can be used to remove the DNase, as well as divalent cations such as magnesium and calcium, which can catalyze RNA degradation when RNA is heated with the sample.

How can I inactivate DNase I?

Add of 1 µL of 25 mM EDTA solution to the reaction mixture in 10 µL reaction with 1 unit DNase I, Amplification Grade (or 1:1 molar ratio of Mg++ ions:EDTA) to chelate the Mg++ ions in the DNase I buffer. Heat for 10 min at 65 degrees C.

Note: It is vital that the EDTA be at least at 2 mM prior to heat-inactivation to avoid Mg-dependent RNA hydrolysis.

DNA-free and Turbo-free versions of DNase I can be inactivated with included DNase Inactivation Reagent.

Are your DNase I products RNase-free?

Most of our DNase I products are guaranteed free of RNase activity. However, please note that Cat. No. 18047-019 is not tested for RNAse and is recommended primarily for protein applications. The other products are suitable for removing DNA from both RNA and protein preparations, for nick translating DNA, and for generating random fragments of DNA. For more demanding RT-PCR applications, we recommend using DNAse I, Amplification Grade.

What are the ideal storage conditions for the TURBO DNase (2 U/μl) (Cat. No. AM2239, AM2238)? Will the enzyme remain stable even after multiple freeze/thaw cycles?

The TURBO DNAse itself is contained in a buffer containing glycerol, and therefore it will not freeze at -20°C. The reaction buffer is an aqueous salt solution which will remain stable even after multiple freeze/thaw cycles. We do not recommend storing it under different conditions. You could also aliquot the DNAse, but under these circumstances it wouldn't be necessary.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

Does TURBO DNase contain DTT?

TURBO DNase does not contain DTT.

Can I purchase the 10X Reaction Buffer included with TURBO DNase as a stand-alone product?

We do not offer the 10X Reaction Buffer that comes with TURBO DNase as a stand-alone item and the composition of the buffer is proprietary.

Does TURBO DNase cleave both single- and double-stranded DNA?

Yes, since the cleavage site for TURBO DNase is the phosphodiester bond, it cleaves both single- and double-stranded DNA.

How does the dsDNase in Maxima First Strand cDNA Synthesis Kit for RT-qPCR, with dsDNase (Cat. Nos. K1671, K1672) compare with TURBO DNase?

The main difference between the two enzymes is that dsDNase is heat-labile (easily inactivated by moderate heat treatment (55 degrees C)), and hence does not need a separate heat inactivation step. It gets inactivated during the RT reaction (cDNA synthesis) itself. Turbo DNase on the other hand is not heat-labile and thus needs to be heat inactivated post-digestion or has to be removed by phenol-chloroform extraction.

I want to use the MagMAX-96 for Microarrays Total RNA Isolation Kit to isolate total RNA with the modified spin protocol (https://bit.ly/2Z9cPQK). Can I add the TURBO DNase steps from the standard no-spin protocol to the modified spin protocol?

When using the MagMAX-96 for Microarrays Total RNA Isolation Kit modified spin protocol (https://www.thermofisher.com/us/en/home/references/protocols/nucleic-acid-purification-and-analysis/rna-protocol/isolating-small-rnaas-using-the-magmax-96-for-microarrays-kit.html) for isolating large + small RNA, we recommend going through the RNA isolation procedure and then treating the eluted RNA with TURBO DNase.