Description

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.

Source

Recombinant E. coli

Concentration

0.1U/μL, 1U/μL (customized)

Application

Homogenization of Full-length cDNA Library

Single nucleotide polymorphism (SNP) detection

high-throughput sequencing

Detection of microRNAs (miRNAs)

Quality

purity>95% by SDS PAGE

Storage

Store at -20°C

Unit Definition

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.

Package

Component	DSN0101	DSN0102	DSN0103
DSN	50U	100U	1000U
10×DSN Reaction Buffer	0.5ml	0.5ml	3ml
2×DSN Stop Buffer	1ml	1ml	6ml

Protocol

1. Prepare the reaction according following table on ice.

Components	Quantity
50-500 ng DNA	x μl
10×DSN Reaction Buffer	1 μ
DSN	y μl
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.

Note

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.

Order

DSN01

Duplex specific nuclease

0.1U/μl Specific nuclease enzyme hydrolyze DNA in dsDNA or DNA-RNA