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Cod UNG

Cod UNG

For diagnostic assays

Cod UNG

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Cod Uracil-DNA Glycosylase (Cod UNG) from Atlantic Cod is the only commercially available UNG enzyme that is completely and irreversibly inactivated by moderate heat treatment. The enzyme is produced in a recombinant E. coli (ung-) strain that contains a modified Cod UNG gene.

Key Features:

  • Heat-labile - Completely and irreversibly inactivated at 55°C
  • Contamination control - ideal in applications below
  • Use of Cod UNG makes contamination control possible in RT-PCR
  • Does not degrade PCR product post-PCR. This makes downstream use of the PCR product possible
  • High purity enzyme, tested free of contaminating nucleases
  • Detergent free
  • Post-PCR Analysis - Enables post-PCR analysis

Applications

Ideal for contamination control in 

Heat-labile Uracil-DNA Glycosylase

There are several commercially available Uracil-DNA glycosylases on the market today. Most of them are of bacterial origin and work well if you have no intention to further analyze the PCR products post-PCR. However, if you want to store your PCR products for downstream analysis such as cloning and sequencing, the reactivation of UNG and subsequent degradation of your PCR products are a problem with most of the commercially available UNGs. Cod UNG from ArcticZymes is completely and irreversibly inactivated by heat thus ensuring that sample integrity is maintained long-term regardless of storage conditions.

This is illustrated in figure 1, below

Figures

Cod UNG - The only UNG that become completely and irreversibly heat-inactivated is Cod UNG
Figure 1. The only UNG that become completely and irreversibly heat-inactivated is Cod UNG
Cod UNG - The only UNG that become completely and irreversibly heat-inactivated is Cod UNG

Various UNGs were tested for residual activity after heat inactivation. PCR was performed with dUTP and 1 Unit of 5 different commercially available UNGs. Post-PCR, the PCR products were incubated at room temperature for various time intervals, followed by heating and subsequent cooling. Gel electrophoreses of the PCR products revealed UNG reactivation, and thus severe degradation of PCR products of all UNGs tested, except for

Cod UNG - Chromatograms of sequenced PCR products pre-treated with 1 U Cod UNG (A) or 1 U E.coli UNG (B) and incubated at room temperature for 3 hours.
Figure 2. Chromatograms of sequenced PCR products pre-treated with 1 U Cod UNG (A) or 1 U E.coli UNG (B) and incubated at room temperature for 3 hours. Only Cod UNG leaves sequence quality intact.
Cod UNG - Chromatograms of sequenced PCR products pre-treated with 1 U Cod UNG (A) or 1 U E.coli UNG (B) and incubated at room temperature for 3 hours.

Post-PCR sequence quality and integrity were further evaluated by sequencing the PCR products. PCR was performed with one of four different commercially available UNGs added to the mastermix. Post-PCR, the PCR products were incubated at room temperature or 4˚C at various time intervals. Samples were subsequently purified and sequenced. Sequence data were thoroughly analyzed with emphasis on reduced sequence quality as a result of UNG reactivation. As illustrated in both figure 2 and figure 3, samples treated with UNG showed severe degradation of PCR products due to UNG reactivation, except for samples treated with Cod UNG.

Cod UNG - Cod Uracil-DNA Glvcosvlase - ArcticZvmes Technologies
Figure 3. Cod Uracil-DNA Glvcosvlase - ArcticZvmes Technologies
Cod UNG - Cod Uracil-DNA Glvcosvlase - ArcticZvmes Technologies

UNG reactivation resulted in degraded sequences. Sequence data Of PCR products pretreated with various LJNGs and incubated at room temperature. All samples, except samples incubated With Cod UNG, demonstrated reactivation of UNG and severe degradation of PCR products.

Cod UNG - Cod UNG restores sensitivity
Figure 4. Cod UNG restores sensitivity
Cod UNG - Cod UNG restores sensitivity

A serial dilution of MS2 viral RNAin human serum was prepared. The mixture was spiked with uracilcontaining amplicons to mimic carry-over DNA. RNA was quantifiedusing a standard one-step RT-qPCR. The presence of carry-overDNA severely reduced the sensitivity of the assay (red). Inclusion ofCod UNG restored assay sensitivity (green), control sample (grey)

Cod UNG - Cod UNG remains inactive
Figure 5. Cod UNG remains inactive
Cod UNG - Cod UNG remains inactive

RT-qPCR on human total RNA in presence of Cod UNG, UNG from E. coli, and two competitor coldadapted UNG’s was performed according to protocol. The failureof competitors A and B (cold-adapted UNG’s from marine microorganisms) to fully inactivate at 50-60°C leads to loss of cDNA andthe integrity of the PCR reaction.

Cod UNG - Cod UNG activity in the presence of blood components
Figure 6. Cod UNG activity in the presence of blood components
Cod UNG - Cod UNG activity in the presence of blood components

Cod UNG has a high tolerance for blood components such asserum and anti-coagulants, including EDTA and Na-Heparin.

Cod UNG - Cod UNG is irreversibly heat-inactivated
Figure 7. Cod UNG is irreversibly heat-inactivated
Cod UNG - Cod UNG is irreversibly heat-inactivated

Cod UNG was testedside-by-side two commercially available, heat-labile UNG enzymes from cold-adapted marine microorganisms in a typical RTbuffer. The enzymes were heat-inactivated at the indicated timeand temperatures. Following inactivation, PCR product was addedto the samples and incubated for three hours at 37°C. Despite inactivation, upon incubation at 37°C, competitor enzymes degradedDNA (faint bands), while Cod UNG remained inactive in all bufferconditions.

Properties

Specifications
Unit Definition

One Unit is defined as the amount of enzyme that liberates 1 nmol Uracil per hour from Uracil-labelled DNA at 37°C in 70 mm Tris-HCI pH 8.0 (@25°C), 10 mM NaCI, 1 mM EDTA and 0.1 mg/ml BSA.

Specific activity

> 500 000 Units/mg

Purity

Purified to apparent homogeneity by SDS-PAGE. No nuclease activity is detected.

Concentration

Minimum 1 000 Units /ml.

Stability

Minimum shelf life for standard formulation at -20°C is 3 years. In practice we find that storage at 4°C is possible for at least 6 months. The enzyme activity is also preserved upon multiple freeze-thaw cycles.

Quality Control

Recommended Protocols

1. Contamination control in PCR, qPCR and one-step RT-qPCR

Cod UNG works in all commercially available master mixes. 

Be sure that you have used dUTP containing dNTP mixes in your previous PCR experiments.

  • Add 0.2 U Cod UNG directly to your 20 µl PCR reaction.
  • pre-incubate for 5 min at room temperature.
  • For RT-qPCR, reverse transcribe your RNA at 50-55°C.
  • Run your PCR.
  • Store your PCR product at -20°C or 4°C degrees.

2. Contamination control in RT-LAMP

Cod UNG is ideal for contamination control in RT-LAMP.  
One unit of Cod UNG per 30 μl reaction is sufficient for removing even high concentrations of carry-over contamination.

  • Ensure that you use dNTP mixes containing dUTP in your experiments.
  • Check that the RT-LAMP reaction is compatible with dUTP by running side-by-side reactions containing different rations of dUTP to dUTP (100% dUTP, 90% dUTP, 80% dUTP and 0% dUTP).
  • Add 1 U Cod UNG directly to your 30 µl RT-LAMP reaction.
  • Prepare the reaction mix on ice.
  • Analyze your RNA at 65°C, no preincubation is necessary.

Please refer to Protocol for Carry-over Contamination Control

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
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
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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
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
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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
Request Quote
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
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
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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
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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
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
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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
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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
9
HL-dsDNase
70800-150
50 kU
50 U/µl
$ 30,330.00 USD
26368.00
30330.00
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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
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10
HL-dsDNase
70800-110
100 kU
2 U/µl
$ 45,552.00 USD
39608.00
45552.00
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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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11
HL-dsDNase
70800-120
500 kU
2 U/µl
$ 170,682.00 USD
148412.00
170682.00
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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
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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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