Recombinant DNase I (power, RNase-free)
Cat No: HC4007B
DNase I is an endonuclease that can digest single-or double-stranded DNA. It can hydrolyze phosphodiester bonds to produce mono-and oligodeoxynucleotides containing a 5'-phosphate group and a 3'-OH group. The optimal working pH range of DNase I 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. In the presence of Mg2+, DNase I can randomly cleave any site of double-stranded DNA; While in the presence of Mn2+, DNase I can cleave DNA double-stranded at the same site, forming blunt ends or sticky ends with 1-2 nucleotides protruding. It can be used for the processing of various RNA samples.
Components
|
Name |
2.5KU |
|
DNaseI, RNase-free |
2.5KU |
|
10× DNase Buffer |
2.5 mL |
Storage conditions
This product should be stored at-25~- 15℃ for 2 years. Please avoid repeated freeze thaw.
Instructions for Use
1. Preparation of DNase I storage solution: Dissolve 2500U of DNase I Lyophilized powder in 500μL RNase-free ddH2O, gently mix, pack and store at -25 ~ -15°C, stable storage for 12 months. Retrieve as needed according to experimental requirements each time. Please note that the DNase I storage solution stored at -25~ -15℃ should not undergo more than five freeze-thaw cycles. Alternatively, store the DNase I storage solution at 2~8℃ (stable storage for 2 months).
2. Prepare the reaction solution in the RNase-free tube according to the proportions listed below:
|
Component |
Volume |
|
RNA |
X µg |
|
10 × DNase I Buffer |
1μL |
|
DNase I, RNase-free(5U/μL) |
1 U per µg RNA① |
|
ddH2O |
Up to 10μL |
1. Calculate the volume of DNase I that needs to be added based on the amount of RNA.
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.
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℃.
Quality Control
RNase Activity: 5U of DNase I with 1.6 µg MS2 RNA for 4 hours at 37℃ yields no degradation as determined by agarose gel electrophoresis.
Notes
1. This product is RNase-free DNase I in powder form. In order to avoid loss in the process of dissolving, please carefully open the bottle cap and slowly add RNase-free ddH2O directly to the bottle; Or please do not open the bottle cap, directly use the RNase-free syringe to inject the RNase-free ddH2O into the bottle, inversely mix until the DNase I is completely dissolved, and then use the RNase-free syringe to suck out the solution (the RNase-free syringe should be prepared by yourself).
2. Do not swirl violently when preparing DNase I storage solution. Mix gently upside down.
3. Please prepare 0.5M EDTA by yourself.
4. Use 1U DNaseI per µg of RNA. However, if the RNA is less than 1 µg, please use 1 U DNase I.
5. Please place the enzyme on ice to avoid repeated freezing and thawing.
