Duplex specific nuclease (DSN) is a specific nuclease enzyme, which has strong activity to hydrolyze DNA in dsDNA or DNA-RNA hybrid duplexes. The DSN has very slight activity for hydrolysis ssDNA or RNA.
Recombinant E. coli
0.1U/μL, 1U/μL (customized)
Homogenization of Full-length cDNA Library
Single nucleotide polymorphism (SNP) detection
Detection of microRNAs (miRNAs)
purity>95% by SDS PAGE
Store at -20°C
Under the standard reaction system, the increase of 0.001 per minute in the absorbance of 50 𝜇g/ml calf thymus DNA at 25 ℃ is defined as 1 unit.
|10×DSN Reaction Buffer||0.5ml||0.5ml||3ml|
|2×DSN Stop Buffer||1ml||1ml||6ml|
1. Prepare the reaction according following table on ice.
|50-500 ng DNA||x μl|
|10×DSN Reaction Buffer||1 μ|
|ddH2O up to||10 μl|
2. Mix gently and centrifugate briefly.
3. Incubate at 65 ℃ for 7-20 min.
4. Add 5 μl 2 × DSN Stop Buffer, gently mixed, centrifuged briefly, 65 ℃, incubated for 5 min to terminate the reaction.
1. DSN digestion time depends on the sample and specific experimental requirements. The incubation temperature can vary from 35 to 70 ℃; In this case, the incubation time and DSN concentration need to be optimized separately.
2. DSN is active in the presence of divalent cations (Mn2+, Co2+ or Mg2+). The concentration of Mg2+ions in most applications should be at least 5 mM. EDTA can inhibit DSN activity.
3. The optimum temperature for DSN activity is 60 ℃. However, DSN retains only 10% activity at 80 ℃.
4. DSN is resistant to protease K treatment (incubation at 37 ℃ for 30 minutes).
5. DSN is highly sensitive to salt ion concentration (for example, the activity decreases by 10 times under 0.2M NaCl.
|DSN01||Duplex specific nuclease||0.1U/ul Specific nuclease enzyme hydrolyze DNA in dsDNA or DNA-RNA|