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Endonucleases Non-Specific

Endonucleases Non-Specific

Endonucleases Non-Specific

Endonucleases Non-Specific, HL-SAN

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HL-SAN efficiently removes nucleic acids from buffers typically used in protein purification. Due to its high salt tolerance, it is the obvious choice for host-cell DNA removal in settings where salt is added to reduce aggregation. Especially efficient for removing nucleic acids from proteins with high affinity for DNA and RNA. Proven performance during lysis and early stages of protein purification processes, as well as high-salt eluates. HL-SAN is a cold-adapted enzyme with excellent performance also at ambient temperatures and during over-night digestion at 4°C.

Recommended applications:

Complete removal of DNA 

Viscosity reduction

Key advantages with HL-SAN:

  • Non specific endonuclease
  • Optimum activity at high salt concentration (0.5 M NaCl)
  • Active at low temperatures (20% at 6ºC)
  • Easily inactivated
  • Broad pH range
  • Temperature stable

Figures

Endonucleases Non-Specific - Optimum activity in solutions with high salinity
Figure 1. Optimum activity in solutions with high salinity
Endonucleases Non-Specific - Optimum activity in solutions with high salinity

HL-SAN has optimum activity at ∼0.5 M NaCl, but operates at a broad range of [NaCl] and [KCl]. The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 5 mM MgCl2 with varying [NaCl] or [KCl]. The maximum activity was set to 100%.

Endonucleases Non-Specific - Temperature and activity
Figure 2. Temperature and activity
Endonucleases Non-Specific - Temperature and activity

HL-SAN has optimum activity at ~35°C, but works over a broad temperature range (20% activity at 10°C and 50°C). The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5 containing 5 mM MgCl2 and 0.5 M NaCl.

Endonucleases Non-Specific - The effect of MgCl2 and MnCl2 concentration on the HL-SAN activity.
Figure 3. The effect of MgCl2 and MnCl2 concentration on the HL-SAN activity.

Fig 3. The effect of MgCl2 and MnCl2 concentration on the HL-SAN activity.

Endonucleases Non-Specific - The effect of MgCl2 and MnCl2 concentration on the HL-SAN activity.

The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 0.5 M NaCl and with varying concentrations of MgCl2 or MnCl2. The activity of the sample containing 5 mM MgCl2 was set to 100%.

Endonucleases Non-Specific - HL-SAN activity vs pH/[NaCl]
Figure 4. HL-SAN activity vs pH/[NaCl]
Endonucleases Non-Specific - HL-SAN activity vs pH/[NaCl]

The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer with different pHs and different concentrations of NaCl. All buffers contained 5 mM MgCl2. The nature of the buffer was pH-dependent, but generally the NaCl-optimum was the same in all buffers/pHs. The exception was etanolaminbuffer at pH 9 and pH 9.5 in which the NaCl-optimum was shifted to the left (not shown).

Endonucleases Non-Specific - Buffer composition affects substrate preference
Figure 5. Buffer composition affects substrate preference
Endonucleases Non-Specific - Buffer composition affects substrate preference

Without NaCl, the specificity towards ssDNA and dsDNA is similar. At 0.5 M NaCl, the activity towards dsDNA increases, while the activity towards ssDNA is unaffected.

Endonucleases Non-Specific - HL-SAN digests ssDNA to ~5-13 nt, and dsDNA to ~5-7 nt
Figure 6. HL-SAN digests ssDNA to ~5-13 nt, and dsDNA to ~5-7 nt
Endonucleases Non-Specific - HL-SAN digests ssDNA to ~5-13 nt, and dsDNA to ~5-7 nt

The size of the end products from ssDNA varies from ~5-13 nt, while dsDNA is digested to around ~5-7 nt. The size of the end products seems to depend on the DNA sequence. Substrates 1 and 2 were ssDNA with different sequences and substrates 3 and 4 were dsDNA with similar sequences but with a FAM-label at different ends. Substrate 5 was dsDNA with the same sequence as substrate 3 and 4 but with a FAM-label at both ends.

Endonucleases Non-Specific - HL-SAN activity decreases with increasing concentrations of glycerol
Figure 7. HL-SAN activity decreases with increasing concentrations of glycerol
Endonucleases Non-Specific - HL-SAN activity decreases with increasing concentrations of glycerol

The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 5 mM MgCl2, 0.5 M NaCl and with increasing concentrations of glycerol. The activity of the control not containing glycerol was set to 100%.

Endonucleases Non-Specific - The activity of HL-SAN at different concentrations of imidazole
Figure 8. The activity of HL-SAN at different concentrations of imidazole
Endonucleases Non-Specific - The activity of HL-SAN at different concentrations of imidazole

The activity of HL-SAN was tested in a 25 mM Tris-HCl buffer, pH 8.5, 5 mM MgCl2, 0.5 M NaCl and with varying concentrations of imidazole. The activity of the control not containing imidazole was set to 100%.

Properties

Specifications
Source

Recombinantly produced in Pichia pastoris.

Working Ranges
  • Temp: 7 – 38°C and 4°C overnight, optimal: 30 – 38°C
  • Salt concentration (NaCI / KCI): 0 - 1000 mM, optimal: 400 - 700 mM
  • Mg2+: >1 mM is required for activity, optimal: 5 – 50 mM
  • pH: 6.8 - 9.5, optimal: 8.0 - 8.9

Note: The working range is defined as above 10% of activity and optimal range as above 80% activity.

Specific Activity

≥ 175 000 Units/mg

Unit Definition

One Unit is defined as an increase in absorbance at 260 nm of 1 A in 30 minutes at 37°C, using 50 μg/ml calf thymus DNA (D-1501, Sigma) in a buffer consisting of 25 mM Tris-HCl, pH 8.5 (25°C), 5 mM MgCl2, 500 mM NaCl.

Tolerance to Typical Lysis Buffer Additives
  • Imidazole: 20% activity at 350 mM Imidazole
  • Glycerol: 20% activity at 35% glycerol
  • Triton X-100: No reduction in activity in Triton X-100 (tested up to 15%)
  • Tween20: No reduction in activity (tested up to 10%)
  • SDS: Not recommended
  • Urea: Not recommended
  • Reducing agent (e.g. DTT, TCEP): will result in inactivation
Quality Control

ArcticZymes is dedicated to the quality of our products. HL-SAN is manufactured at our ISO 13485 certified facility in Norway.

dsDNA Endonuclease Activity

25 U IsoPol® BST+ was incubated with a supercoiled plasmid (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any transformation of closed circular DNA to nicked DNA.

ssDNA Endonuclease Activity

25 U IsoPol® BST+ was incubated with M13 ssDNA (0.5 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of ssDNA degradation.

Recommended Protocols

Ordering Info

Quantity
Product Name
SKU
Pack Size
Concentration
Price
1
T7 RNA Polymerase
74100-201
10 000 U
50 U/µl
$ 145.00 USD
133.00
145.00
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2
T7 RNA Polymerase
74100-110
100 kU
≥ 500 U/µl
$ 0.00 USD
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3
T7 RNA Polymerase
74100-100
As per request
As per request
$ 0.00 USD
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4
T7 RNA Polymerase Glycerol Free
74110-110
100 kU
≥ 500 U/µl
$ 0.00 USD
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5
T7 RNA Polymerase Glycerol Free
74110-100
As per request
As per request
$ 0.00 USD
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1
T4 DNA Ligase
71800-202
250 kU
5000 U/μl
$ 32.00 USD
30.00
32.00
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1
AZscript RT
72100-201
10000 U
200 U/μl
$ 136.00 USD
123.00
136.00
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1
Heat&Run gDNA Removal Kit
80200-250
250 rxn
$ 651.00 USD
548.00
651.00
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1
M-SAN HQ ELISA Kit
70960-001
12x8 Strip plate
$ 756.00 USD
655.00
756.00
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1
HL-dsDNase Triton Free
70801-201
250 U
2 U/µl
$ 357.00 USD
305.00
357.00
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1
PCR Decontamination Kit
80400-100
100 rxn
$ 130.00 USD
110.00
130.00
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1
AZtaq™ DNA Polymerase
73100-201
500 U
5 U/µl
$ 83.00 USD
75.00
83.00
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1
HL-SAN Triton Free
70911-202
25000 U
25 U/µl
$ 560.00 USD
473.00
560.00
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1
ArcticZymes Proteinase
71600-201
50 U
>200 U/ml
$ 155.00 USD
131.00
155.00
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1
IsoPol®
71500-201
200 U
5 U/µl
$ 75.00 USD
68.00
75.00
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1
ArcticZymes R2D Ligase
71900-105
5 kU
10 U/µl
$ 419.00 USD
361.00
419.00
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1
rSAP
70700-201
1000 U
1 U/µl
$ 131.00 USD
116.00
131.00
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1
dsDNase Triton Free
70601-201
250 U
2 U/µl
$ 357.00 USD
305.00
357.00
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1
Cod UNG Triton Free
70501-201
100 U
1 U/µl
$ 130.00 USD
109.00
130.00
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1
IsoPol® BST⁺
71502-202
1600 U
8 U/µl
$ 78.00 USD
71.00
78.00
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1
SAN High Quality
70920-150
500 kU
>250 U/µl
$ 5,767.00 USD
5011.00
5767.00
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1
HL-ExoI
70100-201
2500 U
>20 U/µl
$ 69.00 USD
65.00
69.00
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1
SAN HQ 2.0
70940-150
500 kU
250 U/µl
$ 5,767.00 USD
5011.00
5767.00
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2
Cod UNG Triton Free
70501-202
1000 U
1 U/µl
$ 548.00 USD
456.00
548.00
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2
SAN High Quality
70920-160
5 MU
>250 U/µl
$ 21,913.00 USD
19035.00
21913.00
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2
PCR Decontamination Kit
80400-500
500 rxn
$ 519.00 USD
445.00
519.00
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2
HL-dsDNase Triton Free
70801-202
1000 U
2 U/µl
$ 1,055.00 USD
877.00
1055.00
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2
IsoPol® BST⁺
71502-120
10000 U
120 U/µl
$ 384.00 USD
350.00
384.00
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2
dsDNase Triton Free
70601-202
1000 U
2 U/µl
$ 1,055.00 USD
877.00
1055.00
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2
T4 DNA Ligase
71800-100
as per request
as per request
$ 0.00 USD
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2
ArcticZymes R2D Ligase
71900-100
As per request
As per request
$ 0.00 USD
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2
M-SAN HQ
70950-120
200 kU
>250 U/µl
$ 3,155.00 USD
3024.00
3155.00
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2
IsoPol® SD+
71501-201
200 U
5 U/µl
$ 75.00 USD
68.00
75.00
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2
HL-SAN Triton Free
70911-150
500 kU
25 U/µl
$ 5,425.00 USD
4714.00
5425.00
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2
rSAP
70700-202
5000 U
1 U/µl
$ 519.00 USD
450.00
519.00
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2
SAN HQ 2.0
70940-160
5 MU
>250 U/µl
$ 21,913.00 USD
19035.00
21913.00
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2
Heat&Run gDNA Removal Kit
80200-50
50 rxn
$ 166.00 USD
139.00
166.00
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2
HL-ExoI
70100-202
500 kU
>20 U/µl
$ 6,148.00 USD
5040.00
6148.00
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2
AZscript RT
72100-100
As per request
As per request
$ 0.00 USD
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2
ArcticZymes Proteinase
71600-110
1000 U
>200 U/ml
$ 1,419.00 USD
1188.00
1419.00
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2
AZtaq™ DNA Polymerase
73100-110
5000 U
>50 U/µl
$ 619.00 USD
568.00
619.00
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3
Cod UNG Triton Free
70501-110
10 kU
1 U/µl
$ 3,920.00 USD
3472.00
3920.00
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3
SAN HQ 2.0
70940-202
25 kU
25 - 30 U/µl
$ 577.00 USD
498.00
577.00
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3
rSAP
70700-203
25 kU
1 U/µl
$ 1,965.00 USD
1703.00
1965.00
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3
HL-ExoI
70100-150
50 kU
>20 U/µl
$ 683.00 USD
593.00
683.00
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3
M-SAN HQ
70950-150
500 kU
>250 U/µl
$ 6,055.00 USD
5803.00
6055.00
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3
ArcticZymes Proteinase
71600-100
As per request
>200 U/ml
$ 0.00 USD
Request Quote
3
IsoPol® BST⁺ Glycerol Free
71512-100
As per request
As per request
$ 0.00 USD
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3
HL-dsDNase Triton Free
70801-203
2500 U
5 U/µ
$ 2,365.00 USD
1961.00
2365.00
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3
AZtaq™ DNA Polymerase
73100-100
As per request
As per request
$ 0.00 USD
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3
dsDNase Triton Free
70601-203
2500 U
5 U/µl
$ 2,365.00 USD
1961.00
2365.00
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3
SAN High Quality
70920-202
25 kU
25-30 U/µl
$ 577.00 USD
498.00
577.00
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3
HL-SAN
70910-202
25000 U
25 U/µl
$ 560.00 USD
473.00
560.00
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4
IsoPol® BST⁺ Glycerol HC Free
71522-100
As per request
As per request
$ 0.00 USD
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4
Cod UNG Triton Free
70501-151
50 kU
1 U/µl
$ 14,128.00 USD
12281.00
14128.00
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4
ArcticZymes Proteinase Glycerol-FREE
71610-100
As per request
>200 U/ml
$ 0.00 USD
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4
rSAP
70700-101
200 kU
1 U/µl
$ 13,375.00 USD
11631.00
13375.00
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4
dsDNase Triton Free
70601-150
50 kU
50 U/µl
$ 17,069.00 USD
14830.00
17069.00
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4
SAN HQ 2.0 ELISA Kit
70970-001
1x96 Well Plate
$ 755.00 USD
655.00
755.00
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4
HL-dsDNase Triton Free
70801-110
100 kU
2 U/µl
$ 45,552.00 USD
39608.00
45552.00
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4
HL-SAN
70910-150
500 kU
26 U/µl
$ 5,425.00 USD
4714.00
5425.00
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4
SAN High Quality GMP
70980-150
500 kU
400-600 U/ul
$ 0.00 USD
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5
Cod UNG Triton Free
70501-150
50 kU
50 U/µl
$ 14,128.00 USD
12281.00
14128.00
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5
HL-dsDNase Glycerol-Free - Triton Free
70811-50
5000 U
>50 U/µl
$ 4,031.00 USD
3505.00
4031.00
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5
rSAP Glycerol-free
70710-201
5000 U
>20 U/µl
$ 603.00 USD
521.00
603.00
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4
M-SAN HQ
70950-155
1 MU
>250 U/µl
$ 10,905.00 USD
10450.00
10905.00
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5
M-SAN HQ
70950-160
5 MU
>250 U/µl
$ 23,008.00 USD
22049.00
23008.00
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5
dsDNase Triton Free
70601-110
100 kU
2 U/µl
$ 25,604.00 USD
22260.00
25604.00
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5
SAN High Quality GMP
70980-160
5 MU
400-600 U/µl
$ 0.00 USD
Request Quote
6
rSAP Glycerol-free
70710-100
As per request
As per request
$ 0.00 USD
Request Quote
6
Cod UNG Glycerol-Free Triton Free
70511-105
5000 U
50 U/µl
$ 2,598.00 USD
2256.00
2598.00
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6
dsDNase Glycerol Free, Triton Free
70611-50
5000 U
>50 U/µl
$ 4,031.00 USD
3505.00
4031.00
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6
SAN High Quality Triton Free
70921-150
500 kU
>250 U/µl
$ 5,767.00 USD
5011.00
5767.00
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6
M-SAN HQ
70950-202
25 kU
25-30 U/µl
$ 606.00 USD
580.00
606.00
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6
HL-dsDNase
70800-201
250 U
2 U/µl
$ 357.00 USD
305.00
357.00
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7
Cod UNG Glycerol-Free Triton Free
70511-150
50 kU
50 U/µl
$ 14,128.00 USD
12281.00
14128.00
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7
SAN High Quality Triton Free
70921-160
5 MU
>250 U/µl
$ 21,913.00 USD
19035.00
21913.00
Request Quote
7
dsDNase
70600-201
250
2 U/µl
$ 357.00 USD
305.00
357.00
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7
HL-dsDNase
70800-202
1000 U
2 U/µl
$ 1,055.00 USD
877.00
1055.00
Request Quote
8
SAN High Quality Triton Free
70921-202
25 kU
25-30 U/µl
$ 577.00 USD
498.00
577.00
Request Quote
8
Cod UNG
70500-201
100 U
1 U/µl
$ 130.00 USD
109.00
130.00
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8
dsDNase
70600-202
1 000
2 U/µl
$ 1,055.00 USD
877.00
1055.00
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8
HL-dsDNase
70800-203
2500 U
5 U/µl
$ 2,365.00 USD
1961.00
2365.00
Request Quote
9
Cod UNG
70500-202
1000 U
1 U/µl
$ 548.00 USD
456.00
548.00
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9
SAN High Quality ELISA Kit
70930-001
1x96 Well Compact Plate Format
$ 756.00 USD
655.00
756.00
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9
dsDNase
70600-203
2 500
5 U/µl
$ 2,365.00 USD
1961.00
2365.00
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10
dsDNase
70600-150
50 kU
50 U/µl
$ 17,069.00 USD
14830.00
17069.00
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10
Cod UNG
70500-110
10 kU
1 U/µl
$ 3,920.00 USD
3472.00
3920.00
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11
dsDNase
70600-110
100 kU
2 U/µl
$ 25,604.00 USD
22260.00
25604.00
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11
Cod UNG
70500-151
50 kU
1 U/µl
$ 14,128.00 USD
12281.00
14128.00
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12
Cod UNG
70500-150
50 kU
50 U/µl
$ 14,128.00 USD
12281.00
14128.00
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12
dsDNase Glycerol Free
70610-50
5000 U
80 - 140 U/µl
$ 4,031.00 USD
3505.00
4031.00
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12
HL-dsDNase Glycerol-free
70810-50
5000 U
80 - 140 U/µl
$ 4,031.00 USD
3505.00
4031.00
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13
Cod UNG Glycerol Free
70510-105
5000 U
50 U/µl
$ 2,598.00 USD
2256.00
2598.00
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14
Cod UNG Glycerol Free
70510-150
50 kU
50 U/µl
$ 14,128.00 USD
12281.00
14128.00
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Note: Article no. 71502-202 and 71502-120 are provided with IsoPol® BST+ 10X Buffer and MgSO4 .
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