DNase I (RNase Free)

Product content： Component D665537 1000 U DNaseⅠ（RNase Free）,1 U/µl 1 ml Reaction Buffer（with MgCl2）,10× 1 ml 200 mM EDTA 1 mlProduct Introduction:DNase I is a deoxyribonucleic acid endonuclease that requires a divalent cation and can be used to degrade single or double stranded DNA. The principle is that DNase I hydrolyzes phosphate diester bonds to produce single or oligonucleotides with 5 '- phosphate groups and 3' - OH. Both Mg2+and Mn2+can activate the activity of DNase I, and the concentration of Ca2+directly affects the activity of the enzyme. When Mg2+is present, random incisions can be generated on each single strand of double stranded DNA; In the presence of Mn2+, double stranded DNA can be broken and fragmented. Used for the preparation, reverse transcription, and in vitro transcription experiments of RNA without DNA contamination.Preparation and important precautions before the experiment:1. Because DNase I is often used in DNA digestion experiments that require maintaining RNA integrity, RNase contamination is minimized during enzyme preparation and can be safely used for RNA extraction. However, since this enzyme does not contain RNase inhibitors, it is recommended to be cautious of contamination with exogenous RNase when using it.2.DNase I is greatly affected by shear forces. Before use, the centrifuge tube can be inverted and mixed evenly to avoid vortex oscillation.3. Every processing 1 μ The dosage of g RNA and DNase I should not exceed 1U.Instructions for use：1. Taking the preparation of RNA samples for RT-PCR as an example, a 10 µ l reaction system was configured, as shown in the table below:2.Incubate at 2.37 ℃ for 30 minutes.3. Add 1 µ l 200 mM EDTA and incubate at 65 ℃ for 10 minutes to inactivate DNase I and terminate the reaction.4. The processed RNA can be used for RT-PCR. Specifications and Purity: 1000 U.