What is Turbo DNase inactivation reagent?

Ambion® TURBO DNA-free™ DNase Treatment and Removal Reagents are designed to remove contaminating DNA from RNA preparations, and to subsequently remove the DNase and divalent cations from the sample.

How does Turbo DNase work?

TURBO™ DNase cleaves double-stranded DNA nonspecifically to leave 5′ phosphorylated oligodeoxynucleotides. It has increased affinity for DNA-binding and remains active in the presence of salt. Note: this product is just the enzyme.

How do I turn off Turbo DNase?

Incubate at 37°C for 30 minutes. Inactivate TURBO™ DNase using one of the following methods: (Recommended) Perform a phenol/chloroform extraction. Add EDTA to a final concentration of 15 mM, and heat at 75°C for 10 minutes (this inactivates >99% of TURBO™ DNase).

What is a DNase enzyme and why is it used in these experiments?

DNase I is a versatile enzyme that nonspecifically cleaves DNA to release 5′-phosphorylated di-, tri-, and oligonucleotide products (1). A powerful research tool for DNA manipulations, DNase I is used in a range of molecular biology applications.

How does phenol chloroform RNA extraction work?

Phenol-chloroform based RNA extraction relies on the use of acid guanidinium thiocyanate-phenol-chloroform to promote phase separation of biological mixtures and subsequent selective isolation of molecules of interest [1,2].

How do you get rid of DNase?

The best way to remove DNase I from your reaction is to perform a phenol/chloroform extraction or to use a spin column. You can do the heat inactivation step, but that may not completely remove all of the DNase I, and it could interfere with your downstream applications.

How does DNase effect DNA?

Deoxyribonuclease (DNase) enzymes perform a variety of important cellular roles by degrading DNA via hydrolysis of its phosphodiester backbone. Deoxyribonuclease I (DNase I) enzymes cleave single or double-stranded DNA and require divalent metal ions to hydrolyze DNA yielding 3΄-hydroxyl and 5΄-phosphorylated products.

What does DNase break DNA into?

What does DNase destroy?

Deoxyribonuclease I (DNase I) is an endonuclease which is secreted to cleave DNA in the extracellular space down to an average of tetranucleotides with 5′ monophosphate and 3′ hydroxyl DNA ends (Baranovskii, Buneva, & Nevinsky, 2004). Both single-stranded DNA and double-stranded DNA are degraded by DNase I.