Advantages at a glance
- Activity: >3000 units/mg
- MW: appr. 31 kDa
- Appearance: salt-free, freeze-dried powder
- Free shipping
- Shipping within 24h
- 30 days money back guarantee
- Purchase comfortable on account
Product information "DNase I (EC 3.1.21.1)"
DNase I is an endonuclease isolated from bovine pancreas that digests double- and single-stranded DNA into oligo- and mono-nucleotides.
Description
DNAse I is used, among other things, to avoid unwanted cell clumping during tissue disaggregation, which makes dispersion of individual cells difficult. Since tissue disaggregation and subsequent isolation of single cells is always accompanied by rupture and lysis of some cells, DNA is released from these cells into the surrounding medium. The released DNA in the medium is thought to be responsible for undesirable cell clumping during tissue dissociation procedures. This clumping effect can be avoided by adding deoxyribonuclease (DNase) to the dissociation medium and has been described for a variety of different cell and tissue types.
DNase I is suitable for eliminating DNA from RNA preparations prior to sensitive applications, such as RT-PCR. Since no RNA purification procedure removes 100% of the DNA, RNA samples should be digested with DNase I before RT-PCR. A simple 15 minute digestion at room temperature removes the contaminating DNA. DNase I is inactivated by adding the stop solution and heating. Heating also denatures the RNA, so the RNA can be used directly for reverse transcription.
This DNAse I is not RNase-free and has to be chemically treated if used for RNA isolation procedures.
Specifications:
Activity: >3000 units/mg
Appearance: salt-free, freeze-dried powder, chromatographically prepared
MW: appr. 31 kDa
Application:
removes DNA for RNA purification procedures. removes DNA in single cell isolation proceduresSource:
bovine pancreasClassification:
Documents:
CertificateProduct description
The normal DNAse reaction buffer is:
10X buffer:
100mM Tris, pH7.5-8.0
25mM MgCl2
5mM CaCl2
Optional: 50-100mM NaCl
as 1X buffer:
10mM Tris, pH7
2.5mM MgCl2
0.5mM CaCl2
Optional: 5-10mM NaCl
A possible alternative is the following 10X buffer:
200mM Tris, pH8.4
20mM MgCl2
500mM KCl
---------------------------------------------------------------------------------------------------
Inactivation of DNAse I
You can deactivate DNAse I by EDTA. The amount of EDTA depends on the amount/concentration of Mg2+ and or Ca2+ ions in the solution. As a rule of thumb you should use Mg2+/Ca2+:EDTA in a 1:1 molar ratio. E.g. if you use the buffer composition above (2.5mM MgCl2 plus 0.5mM CaCl2) you should use 3mM EDTA to inhibit the DNAse activity.
You can also use an even higher amount of EDTA (about 10% higher), especially if there are other divalent cations in the solution, but you have to be careful if you want to proceed with a reverse transcription after the DNAse digest. As EDTA inhibits also the reverse transcriptase it is not recommended to add to much EDTA for the inhibition of DNAse.
=> For this the preferred method to inhibit DNAse is still the combination of heat and EDTA. In this case you can use not more EDTA than in a 1:1 ratio.
Accessory Items
Overview:Proteinase K is a robust and versatile serine protease used extensively in molecular biology for the digestion of proteins in nucleic acid preparations. This solution, at a concentration of 20 mg/mL, is ideal for a wide range of applications including the preparation of DNA and RNA samples. The PCR grade quality ensures that the enzyme is free from contaminants that could interfere with downstream applications, making it highly suitable for sensitive techniques such as PCR.Product Details:ready-to-useConcentration: 20 mg/mLactive over a wide range of reaction conditionsAn elevation of the reaction temperature from 37°C to 50 - 60°C may increase the activity several timesQuality: PCR grade, ensuring high purity and no interference with PCR reactions.Applications: Suitable for DNA and RNA extractionRemoval of protein contaminantsPreparation of samples for various molecular biology techniques.Benefits:High Purity: The PCR grade quality guarantees minimal contamination, ensuring reliable results in sensitive applications. Versatility: Ideal for various molecular biology procedures, including sample preparation for PCR.Convenience: Ready-to-use solution at an optimal concentration for ease of use. Notes:The recommended working concentration of Proteinase K is 0.05 to 1 mg/mL. The activity of the enzyme is stimulated by 0.2 to 1% SDS and also by 1 to 4M ureaCa2+ protects Proteinase K against autolysis and increases the thermal stabilityStable over a wide pH range: 4.0 to 12.5, optimum pH 7.5 to 8.0Activity optimum: 50°C to 55°CRapid denaturation of enzyme occurs at temperatures above 65°CFeatures:No detected exonuclease, endonuclease or RNase activitySpecific activity: >40 units/mg protein Activity: ≥30 units/mg lyophilizateSolubility: ≥20 mg/mLDNA content ≤10 pg/mgShipping conditions: Shipment on wet iceInhibitors of proteinase KWhich substances can inhibit proteinase K? Proteinase K can be inhibited by a variety of substances. These include ‘denaturing substances’, detergents and proteinase inhibitors. Sensitivity to inhibitors depends pretty much on the specific application (exact pH, concentration, temperature). Inhibitors frequently used in biochemistry and molecular biology are:SDS: High concentrations of SDS can denature and inactivate proteinase K.EDTA: EDTA is a chelating agent that can bind to metal ions that are essential for proteinase K activity.Urea: High concentrations of urea can denature and thereby inactivate proteinase K.Detergents: Some detergents, such as Triton X-100 or Tween® 20, can inhibit proteinase K, especially when the detergents are present in high concentrations.Protease inhibitors: Protease inhibitors, such as phenylmethylsulfonyl fluoride (PMSF), can irreversibly inhibit proteinase K.Chaotropic salts: Depending on the DNA extraction protocol, the presence of chaotropic salts or detergents may increase proteinase K activity. However, the same reagents may also inhibit proteinase K activity.Proteinase K can be obtained from Genaxxon as Powder (M3036) > or as 20mg/mL Solution (M3037) >.Produced by Genaxxon bioscience GmbH, founded in 2002 by Dr. Norbert Tröndle to provide reliable products for PCR and custom cell culture media formulations, the Proteinase K Solution 20 mg/mL offers a dependable and efficient solution for your molecular biology needs. Achieve consistent, high-quality results with an enzyme designed specifically for your requirements.