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Isothermal Low Temperature Range

Isothermal Low Temperature Range

DNA polymerases for point-of-care diagnostic assays

Isothermal Low Temperature Range

Isothermal Low Temperature Range

OverView

To accommodate the need for high quality enzymes for isothermal amplification ArcticZymes developed the IsoPol® series of polymerases. For isothermal amplification at lower temperatures (37°C and below) we offer two enzymes, IsoPol® DNA Polymerase and IsoPol® SD+. Both enzymes exhibit excellent processivity and high strand displacement activity, having 5’-3’ polymerase activity while lacking both 3’-5’and 5’-3’ exonuclease activity.

IsoPol® SD+ is an engineered version of IsoPol® DNA Polymerase with even stronger strand displacement and higher salt tolerance.

Temperature
Range

Optimal
Temperature

Strand
displacement
with salt

Salt tolerance
(NaCI/KCL)
(mM)

Suggested applications

IsoPol® SD+

20 - 42°C

37°C

++++

100 - 350

MDA/SDA, RPA, HAD

IsoPol® DNA Polymerase

20 - 42°C

37°C

++

25 - 110

MDA/SDA, RPA, HAD

Figures

Isothermal Low Temperature Range - Temperature optimum of IsoPol® SD+
Figure 1. Temperature optimum of IsoPol® SD+

Figure 1. Temperature optimum of IsoPol® SD+

Isothermal Low Temperature Range - Temperature optimum of IsoPol® SD+

The enzyme was assayed at the indicated temperatures in 10 mM Tris-HCl pH 8.5, 100 mM NaCl, 5 mM MgCl2, 700 μg/mL denatured herring sperm DNA, 33 μM dNTPs (each), 0.125 μM [3H]-dTTP, 0.8 mM DTT, 0.02% Triton X-100. Samples were incubated for 1 and 5 minutes. Activity was determined measuring radioactive decay [CPM] of incorporated [3H]-dTTPs in acid insoluble material. ΔCPM/min between time point one and two were used as measure of activity. Results are depicted as relative to maximum. The dashed line represents a stipulation of curve progression based on IsoPol® DNA Polymerase

Isothermal Low Temperature Range - Polymerase activity of IsoPol® DNA Polymerase and IsoPol® SD+
Figure 2. Polymerase activity of IsoPol® DNA Polymerase and IsoPol® SD+

Figure 2. Polymerase activity of IsoPol® DNA Polymerase and IsoPol® SD+

Isothermal Low Temperature Range - Polymerase activity of IsoPol® DNA Polymerase and IsoPol® SD+

as function of NaCl concentrations in 10 mM Tris-HCl pH 8.5 (@ 25°C), 5 mM MgCl2, 0.72 µg/µl denatured herring-sperm DNA, 33 µM dNTPs (incl. [3H]-dTTPs), 0.4 µg/ml IsoPol or IsoPol II (6.5 nM). Incubation temperature/time: 37°C/10 min. Activity determination by counting decay events of incorporated [3H]-dTTPs in acid insoluble DNA product in counts per minute [CPM]. Data represented relative to highest activity of each polymerase. Error bars represent 1x standard deviation of biological replicates.

Isothermal Low Temperature Range - Relative activity of IsoPol® DNA Polymerase compared to other commercially available polymerases at 37°C
Figure 3. Relative activity of IsoPol® DNA Polymerase compared to other commercially available polymerases at 37°C
Isothermal Low Temperature Range - Relative activity of IsoPol® DNA Polymerase compared to other commercially available polymerases at 37°C

The enzymes were assayed in 10 mM Tris-HCl pH 8.5, 100 mM NaCl, 5 mM MgCl2 , 700 µg/mL denatured herring sperm DNA, 33 µM dNTPs (each), 0.125 µM [3H]-dTTP, 0.8 mM DTT, 0.02% Triton X-100. Activity of Phi29 polymerase was close to non-detectable in this setup.

Properties

Isopol®

IsoPol® SD+

Strand Displacement Activity

IsoPol® DNA Polymerase exhibits similar strand-displacement activity as that of Bsu DNA polymerase I at 25°C.

IsoPol® SD+ exhibits stronger strand-displacement activity than other polymerases at 25°C and 37°C.

Optimal Reaction Conditions

Optimal performance is achieved in a pH 8.5 buffer supplemented with 5 mM MgCl2 and 25-100 mM salt (NaCl or KCl), at 37°C.

Optimal performance is achieved in a pH 8.5 buffer supplemented with 4-6 mM MgCl2 and 100-350 mM salt (NaCl or KCl), at 37°C.

Inactivation

Temperature > 50°C

Temperature > 50°C

Specifications

Isopol®

IsoPol® SD+

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C.

One unit is defined as the amount of enzyme that will incorporate 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C.

Specific Activity

Min. 15 000 Units/mg

Min. 7 500 Units/mg

Storage & Stability

The enzyme is stable at -20°C for up to 1 year in the supplied storage buffer. The enzyme tolerates at least five freeze-thaw cycles (-80°C) without loss of activity.

The enzyme is stable at -20°C for up to 3 years in the supplied storage buffer. The enzyme tolerates at least five freeze-thaw cycles (-80°C) without loss of activity.

Quality Control

ArcticZymes is dedicated to the quality of our products. IsoPol® polymerases are is manufactured at our ISO 13485 certified facility in Norway.

Isopol®

IsoPol® SD+

dsDNA Endonuclease Activity

25 U IsoPol® DNA Polymerase 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.

25 U IsoPol® SD+ 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® DNA Polymerase 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.

25 U IsoPol® SD+ 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
Request Quote
5
T7 RNA Polymerase Glycerol Free
74110-100
As per request
As per request
$ 0.00 USD
Request Quote
1
T4 DNA Ligase
71800-202
250 kU
5000 U/μl
$ 32.00 USD
30.00
32.00
Request Quote
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
Request Quote
2
ArcticZymes R2D Ligase
71900-100
As per request
As per request
$ 0.00 USD
Request Quote
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
Request Quote
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
Request Quote
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
Request Quote
2
AZscript RT
72100-100
As per request
As per request
$ 0.00 USD
Request Quote
2
ArcticZymes Proteinase
71600-110
1000 U
>200 U/ml
$ 1,419.00 USD
1188.00
1419.00
Request Quote
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
Request Quote
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
Request Quote
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
Request Quote
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
Request Quote
3
HL-SAN
70910-202
25000 U
25 U/µl
$ 560.00 USD
473.00
560.00
Request Quote
4
IsoPol® BST⁺ Glycerol HC Free
71522-100
As per request
As per request
$ 0.00 USD
Request Quote
4
Cod UNG Triton Free
70501-151
50 kU
1 U/µl
$ 14,128.00 USD
12281.00
14128.00
Request Quote
4
ArcticZymes Proteinase Glycerol-FREE
71610-100
As per request
>200 U/ml
$ 0.00 USD
Request Quote
4
rSAP
70700-101
200 kU
1 U/µl
$ 13,375.00 USD
11631.00
13375.00
Request Quote
4
dsDNase Triton Free
70601-150
50 kU
50 U/µl
$ 17,069.00 USD
14830.00
17069.00
Request Quote
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
Request Quote
4
SAN High Quality GMP
70980-150
500 kU
400-600 U/ul
$ 0.00 USD
Request Quote
5
Cod UNG Triton Free
70501-150
50 kU
50 U/µl
$ 14,128.00 USD
12281.00
14128.00
Request Quote
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
Request Quote
7
Cod UNG Glycerol-Free Triton Free
70511-150
50 kU
50 U/µl
$ 14,128.00 USD
12281.00
14128.00
Request Quote
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
Request Quote
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
Request Quote
8
dsDNase
70600-202
1 000
2 U/µl
$ 1,055.00 USD
877.00
1055.00
Request Quote
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
Request Quote
9
SAN High Quality ELISA Kit
70930-001
1x96 Well Compact Plate Format
$ 756.00 USD
655.00
756.00
Request Quote
9
dsDNase
70600-203
2 500
5 U/µl
$ 2,365.00 USD
1961.00
2365.00
Request Quote
10
dsDNase
70600-150
50 kU
50 U/µl
$ 17,069.00 USD
14830.00
17069.00
Request Quote
10
Cod UNG
70500-110
10 kU
1 U/µl
$ 3,920.00 USD
3472.00
3920.00
Request Quote
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
Request Quote
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
Request Quote
Note: Article no. 71502-202 and 71502-120 are provided with IsoPol® BST+ 10X Buffer and MgSO4 .
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FAQ's

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Compatibility with buffer formulations?
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Isothermal Low Temperature Range
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