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Bioprocessing with Salt Active Nucleases  - Physiological Conditions

Bioprocessing with Salt Active Nucleases - Physiological Conditions

No Other EndonucleasePerforms Better at Physiological Conditions

Bioprocessing with Salt Active Nucleases - Physiological Conditions

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Applications

M-SAN HQ  - Peak performance where it matters most at physiological conditions

Medium-Salt Active Nuclease High Quality (M-SAN HQ) is a Bioprocessing Grade nuclease developed for removal of both single and double-stranded DNA and RNA at the physiological salt conditions most often used in bioprocessing and biomanufacturing workflows. M-SAN HQ allows you to directly replace Benzonase without changing your workflow.

This novel, nonspecific endonuclease is active over a broad pH range and displays optimum activity at salt concentrations between 125 – 250 mM. Due to its excellent performance at physiological conditions, M-SAN HQ can be used directly in the cell medium or the harvested supernatant without buffer adjustments. This makes M-SAN HQ ideal for the manufacturing of fragile vectors such as lentiviruses and retroviruses.

M-SAN HQ can be directly used in medium without buffer adjustments The high activity of M-SAN HQ at standard cell medium conditions leads to improved DNA clearance compared to other commonly used nucleases. In the data (see Figure 6), an over 2-fold reduction in residual DNA was achieved. 

Key Benefits

  • Compatibility: Ideal for working with both fragile and robust viral vectors and proteins in a variety of cell media
  • Optimum activity: Optimization for cell media salinity allows both shorter DNA fragments and reduced incubation times.
  • Process Efficiency: Smaller DNA fragments simplifies DSP workflows.
  • Cost-Effectiveness: Reduced need for additional reagents and steps can lower production costs
  • Quality: Maintains the integrity of sensitive or labile biological molecules, ensuring a higher quality end product
  • Time-Saving: Streamlined protocol speeds up the manufacturing process
  • Flexibility: Can be used directly in a variety of media without the need for customization

Key Features 

  • High purity (≥ 99%)
  • No protease detected
  • Endotoxin-tested
  • Animal origin-free production
  • Supplied with extended product documentation

Adapted to use in medium without salinity adjustments 

Optimal activity at physiological salinity and pH makes it ideal for DNA removal from mammalian cell media. (see Figures)

The high activity of M-SAN HQ at the physiological salinity and pH found in standard cell medium conditions leads to improved DNA clearance compared to commonly used nucleases.

Simple to Optimize in Cell Media

M-SAN HQ is uniquely formulated to excel in physiological pH conditions, offering high performance at the commonly used cell media pH of 7.4.

While other nucleases often require more alkaline environments, M-SAN HQ stands out for its adaptability and effectiveness in cell media. It's the nuclease that truly aligns with your bioprocessing needs. (See Figure section)

M-SAN HQ ELISA Kit

The M-SAN HQ ELISA Kit confirms the removal of M-SAN High Quality in bioprocessing and biomanufacturing applications with high accuracy: 

  • 100% (±10%) Measuring range 0.47 – 7.50 ng/ml
  • 100% (±20%) Measuring range 0.12 – 0.47 ng/ml

Features:

  • Fast: 1.5 – 2 hours
  • Sensitive - Quantification range: 0.12 – 7.5 ng/ml
  • Accurate
  • Flexible - 96-well plate in 8-well strip format

Figures

M-SAN HQ (Physiological Conditions) - Optimum activity in solutions at physiological salinity
Figure 1. Optimum activity in solutions at physiological salinity
M-SAN HQ (Physiological Conditions) - Optimum activity in solutions at physiological salinity

M-SAN High Quality has optimum activity between 125 - 250 mM NaCl, which correlates to the salt content in buffers with physiological salinity. The activity was measured at 37°C in a 25 mM Tris-HCl buffer, pH 7.4, 2.5 mM MgCl2 and varying concentrations of NaCl. Maximum activity was set to 100%.

M-SAN HQ (Physiological Conditions) - Outperforms commonly used nucleases at physiological salinity
Figure 2. Outperforms commonly used nucleases at physiological salinity
M-SAN HQ (Physiological Conditions) - Outperforms commonly used nucleases at physiological salinity

Physiological salinity is equivalent to approx. 150 mM NaCl. At this concentration of salt, M-SAN HQ is up to five-fold more efficient than commonly used nucleases with optimum performance in absence of salt. The activity was measured at 37°C in a 25 mM Tris-HCl buffer, pH 7.2, 5 mM MgCl2 and varying concentrations of NaCl. The same number of units of each nuclease was used.

M-SAN HQ (Physiological Conditions) - Highly compatible with physiological pH
Figure 3. Highly compatible with physiological pH
M-SAN HQ (Physiological Conditions) - Highly compatible with physiological pH

M-SAN HQ has optimal performance in the 7.2 - 8.7 pH range. This allows very high performance in most biological buffers. With a working range starting at pH 6.5, M-SAN HQ is also a candidate for DNA removal from insect cell media. The activity was measured at 37°C in a 25 mM Tris-HCl or Bis-Tris buffer at various pH, 2.5 mM MgCl2 and 150 mM NaCl. Activity measured at pH 7.7 was set to 100%.

M-SAN HQ (Physiological Conditions) - Superior performance at 37°C and above
Figure 4. Superior performance at 37°C and above
M-SAN HQ (Physiological Conditions) - Superior performance at 37°C and above

M-SAN HQ is highly efficient for digesting DNA at 37°C, and tolerate temperatures up to 50°C. The activity was measured in a 25 mM Tris-HCl buffer at pH 7.8 (@measurement temperature), 5 mM MgCl2 and 150 mM of NaCl. 37°C was set to 100% as this is the temperature used for defining the activity in Units.

M-SAN HQ (Physiological Conditions) - Efficient Residual DNA Fragmentation
Figure 5. Efficient Residual DNA Fragmentation
M-SAN HQ (Physiological Conditions) - Efficient Residual DNA Fragmentation

Gel analysis showing that  M-SAN HQ effectively reduces residual host cell DNA to under 8 nucleotides.

M-SAN HQ (Physiological Conditions) - M-SAN HQ outperforms other nucleases in removal of host-cell DNA from HEK 293 cells directly in cell media
Figure 6. M-SAN HQ outperforms other nucleases in removal of host-cell DNA from HEK 293 cells directly in cell media
M-SAN HQ (Physiological Conditions) - M-SAN HQ outperforms other nucleases in removal of host-cell DNA from HEK 293 cells directly in cell media

M-SAN High Quality has optimal activity around physiological conditions, which makes it ideal for DNA removal directly in mammalian cell media In this case, HEK 293 cells were grown in DMEM for 48 hrs before nuclease treatment directly in medium (75 U/ml nuclease). The medium was supplemented with Mg2+. to 5 mM. Remaining DNA after 1 hr incubation at 37°C was quantified using Quant-iT™ PicoGreen™ dsDNA Assay Kit.

Properties

Specifications
Source

Recombinantly produced in Pichia pastoris

Molecular weight

24.5 kDa

Protein Purity

> 99% by SDS-PAGE analysis

Isoelectric point

8.62

Unit Definition

One unit is defined as the amount of enzyme that causes a ΔA260 = 1.0 in 30 minutes at 37° in 25 mM Tris-HCl pH 7.6 (@25°C), 2.5 mM MgCl2, 150 mM NaCl, and 50 μg/ml calf thymus DNA.

Specificity

Nonspecific endonuclease cleaving single and double stranded DNA and RNA.

Working ranges
  • Temperature: 20 – 50°C, optimal: 36 – 50°C
  • Salt concentrations (NaCI): 10 – 500 mM, optimal: 125 mM – 250 mM
  • Mg2+: > 0.5 mM is required for activity, optimal: 4 – 15 mM
  • pH: 6.5 – 9.5, optimal: 7.2 – 8.7

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

Tolerance to typical buffer additives
  • DTT and other reducing agents may inactivate M-SAN HQ
  • Urea: Not recommended
  • EDTA: Not recommended
Quality Control

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
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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
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6
rSAP Glycerol-free
70710-100
As per request
As per request
$ 0.00 USD
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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
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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
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8
SAN High Quality Triton Free
70921-202
25 kU
25-30 U/µl
$ 577.00 USD
498.00
577.00
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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
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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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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
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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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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
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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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Bioprocessing with Salt Active Nucleases  - Physiological Conditions
SAN HQ
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