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Endonucleases DNA-specific

Endonucleases DNA-specific

HL-dsDNase for removal of gDNA from RNA preparations

Endonucleases DNA-specific

Endonucleases DNA-specific, HL-dsDNase

OverView

HL-dsDNase is especially developed to remove contaminating genomic DNA from RNA preparations. Figure 1 shows that HL-dsDNase can reduce 50 ng of human gDNA to levels non-detectable by qPCR. In figure 2, a human  total RNA sample was treated with HL-dsDNase and analysed on the Bio-Rad Experion™ System. The results  indicate that HL-dsDNase has minimal impact on RNA quality and quantity.

HL-dsDNase is an engineered version of dsDNase that is rapidly and completely inactivated by incubation for 5 minutes at 58°C at pH 8.0 or above. Chemical inactivation in downstream compatible RT or PCR buffers allows milder heat inactivation and, in some cases, skipping heat inactivation altogether. This makes HL-dsDNase very useful for removal of DNA from RNA preparations since the enzyme may be inactivated using various strategies with reduced risk of auto-degradation of  RNA in the presence of magnesium, making HL-dsDNase an ideal enzyme when working with small volumens of RNA.

HL-dsDNase is also an excellent choice for removal of unwanted external DNA from samples prior to analysis of nucleic acids protected by biological membranes, e.g., bacteria, viruses, and sperm. Especially if using downstream analysis methods that might be affected by host cell DNA, as metagenomic sequencing and STR-profiling. The easy inactivation of HL-dsDNase makes it a fast and efficient alternative to methods as differential extraction, where sample material is often lost.

Recommended applications:

Key advantages with this Salt Active Nuclease:

  • Double-strand DNA specific endonuclease
  • Easily heat-inactivated by very moderate heat treatment
  • High specific activity
  • Useful for removal of DNA from RNA preps

Figures

Endonucleases DNA-specific - Figure 1
Figure 1. HL-dsDNase effectively removes gDNA
Endonucleases DNA-specific - Figure 1

HL-dsDNase removes at least 50 ng of gDNA in a 10 µl reaction volume at pH 7.5.

Endonucleases DNA-specific - Figure 2
Figure 2. HL-dsDNase treatment has minimal impact on RNA integrity
Endonucleases DNA-specific - Figure 2

RNA incubated in buffer (lanes 1-4), water (lanes 5-8) or 0.1 U/µl HL-dsDNase (lane 9-12). Samples were analysed using Bio-Rad Experion™ RNA Eukaryote Total RNA StdSens Assay.

Endonucleases DNA-specific - Figure 3
Figure 3. HL-dsDNase has minimal impact on RNA quantity
Endonucleases DNA-specific - Figure 3

HL-dsDNase effectively removes gDNA from RNA without affecting SDHA (mRNA) quantification.

Properties

Specifications
Unit Definition

One unit is defined as an increase in absorbance at 260 nm of 0.001 per minute, using 50 µg/ml of high MW DNA in 100 mM Na-acetate pH 5.0 and 5 mM MgCl2.

Recommended Reaction and Conditions
  • Temp.:20 - 40°C, optimal: 37°C
  • Salt conc.: (NaCI/KCI): 0 - 50 mM, optimal: 0 mM
  • Mg2+: 4 - 10 mM, minimum: 2.5 mM
  • pH 7-8.3, optimal: 7.5
Storage buffer

20 mM Tris-HCl pH 7.5, 2 mM MgCl2, 10 mM NaCl, 0.01% (v/v) Tween 20, 50% (v/v) glycerol*

*Glycerol-Free version do not contain glycerol.

Purity

HL-dsDNase is purified to apparent homogeneity.

Sample requirement

Purification of nucleic acids prior to HL-dsDNase treatment.

Storage

50% Glycerol: Minimum shelf life 2 years at -20°C.

Glycerol-free: Minimum shelf life 1 year at -20°C.

Both versions tolerates minimum 6 freeze-thaw cycles.

Quality Control

Recommended Protocols

Ordering Info

Quantity
Product Name
SKU
Pack Size
Concentration
Price
1
72100-201
AZscript RT
72100-201
10000 U
200 U/μl
$ 119.00 USD
Request Quote
1
80200-250
Heat&Run gDNA Removal Kit
80200-250
250 rxn
$ 631.00 USD
531.00
631.00
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1
80400-100
PCR Decontamination Kit
80400-100
100 rxn
$ 107.00 USD
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1
71600-201
ArcticZymes Proteinase
71600-201
50 U
>200 U/ml
$ 150.00 USD
127.00
150.00
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2
80400-500
PCR Decontamination Kit
80400-500
500 rxn
$ 431.00 USD
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2
80200-50
IsoPol™ BST⁺
80200-50
50 rxn
$ 161.00 USD
135.00
161.00
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2
72100-100
AZscript RT
72100-100
As per request
As per request
$ 0.00 USD
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2
71600-110
ArcticZymes Proteinase
71600-110
1000 U
>200 U/ml
$ 1,375.00 USD
1151.00
1375.00
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3
71600-100
ArcticZymes Proteinase
71600-100
As per request
>200 U/ml
$ 0.00 USD
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4
71610-100
ArcticZymes Proteinase Glycerol-FREE
71610-100
As per request
>200 U/ml
$ 0.00 USD
Request Quote
Note: Article no. 71502-202 and 71502-120 are provided with IsoPol® BST+ 10X Buffer and MgSO4 .
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