Recombinant DNase I (RNase-free) HYC117
DNase I is an endonuclease that can digest single-stranded and double-stranded DNA to produce single deoxynucleotides or single-stranded or double-stranded oligodeoxynucleotides.It can hydrolyze the phosphodiester bond to produce monodeoxynucleotides and oligodeoxynucleotides containing 5'-phosphate groups and 3'-OH groups. The average digestion product is the smallest polytetranucleotide. DNase I can catalyze many forms of DNA, such as single-stranded DNA,double-stranded DNA, and even chromatin(its cutting rate is affected by histones).The optimum pH range is 7-8. The activity of DNase I depends on Ca²+ and can be activated by divalent metal ions,such as Co2+, Mn²+, Zn²+, etc. 5mM Ca²+ can protect the enzyme from being hydrolyzed. In the presence of Mg²+, the enzyme can recognize and cut any site on any strand of DNA randomly;and in the presence of Mn²+, it can recognize two strands of DNA at the same time and cut at almost the same site to form blunt ends, or sticky ends with 1-2 nucleotides protruding.
Components
|
Components |
1KU |
5KU |
50KU |
|
Recombinant DNase I (RNase-free) |
200μL |
1mL |
10mL |
|
10 × DNase I Buffer |
1mL |
5×1mL |
5×10mL |
Storage
Store at-15°C ~-25℃ for one year.
Unit Definition
One unit is defined as the amount of enzyme which will completely degrade 1 µg of pBR322 DNA in 10 minutes at 37°C.
Specification
|
Source |
Bovine Pancreas DNase I was expressed in yeast expression system and purified. |
|
Storage Buffer |
10 mM Tris-HCl, 2 mM CaCl2, 50% glycerol, pH 7.6 @25℃. |
|
RNase Activity |
5 U of DNase I with 1.6 μg MS2 RNA for 4 hours at 37 ℃ yields no degradation as determined by agarose gel electrophoresis. |
Instructions
1. Prepare the reaction solution in the RNase-free tube according to the proportions listed below
|
Components |
Volume |
|
RNA |
X μg |
|
10×DNase I Buffer |
1 uL |
|
Recombinant DNase I (RNase-free) |
1 U per μg RNA* |
|
RNase-free ddH₂O |
Up to 10μL |
*Calculate the volume of DNase I that needs to be added based on the amount of RNA.Use 1 U DNase I per µg of RNA. However, if the RNA is less than 1 µg, please use 1 U DNase I.
2. 37 ℃ for 15 minutes;
3. Add 0.5M EDTA to the final concentration of 2.5mM~5mM, and heat at 65℃ for 10 minutes to stop the reaction. The sample can be directly used for the next reaction such as reverse transcription experiment.