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T4 DNA Ligase

T4 DNA Ligase

for Ligation of dsDNA, NGS library prep and molecular cloning

T4 DNA Ligase

OverView

T4 DNA Ligase is an ATP and Mg2+ dependent dsDNA ligase which catalyses the formation of a phosphodiester bond between 3’-hydroxyl and 5’-phosphate termini in duplex DNA, duplex RNA and some DNA/RNA hybrids. T4 DNA Ligase is active on both blunt-end and cohesive-end substrates. It is also completely inactivated by incubating at 70°C for 10 minutes.

This is a high-quality (commercial grade) version of the T4 DNA Ligase. T4 DNA Ligase is recombinantly produced in E. coli. ArcticZymes’ T4 DNA Ligase is extensively tested for contaminating DNase and RNase activities as well as residual host-cell gDNA.

Key Features

  • ATP and Mg2+ dependent dsDNA ligase
  • Easily heat-inactivated at 70°C for 10 minutes
  • Extensively tested for contaminating DNase and RNase activities as well
    as residual host-cell gDNA

Applications

  • Ligation of dsDNA
  • NGS library prep
  • Molecular cloning

Figures

T4 DNA Ligase - No residuals of E.coli gDNA were detected in ArcticZymesT4 DNA Ligase
Figure 1. No residuals of E.coli gDNA were detected in ArcticZymesT4 DNA Ligase
T4 DNA Ligase - No residuals of E.coli gDNA were detected in ArcticZymesT4 DNA Ligase

T4 DNA Ligase activity on various substrates. In vitro assays of ligation activity were performed using three oligos hybridised into a nicked 20 bp dsDNA substrate. Efficient DNA ligation was observed after 30 min at 25°C when using 1.12 U/μl T4 DNA Ligase. At 0.373 U/μl T4 DNA Ligase showed about 70% substrate turnover.

T4 DNA Ligase -T4 DNA Ligase displays good nick-joining activity of dsDNA
Figure 2. T4 DNA Ligase displays good nick-joining activity of dsDNA
T4 DNA Ligase -T4 DNA Ligase displays good nick-joining activity of dsDNA

T4 DNA Ligase activity on various substrates. In vitro assays of ligation activity were performed using three oligos hybridised into a nicked 20 bp dsDNA substrate. Efficient DNA ligation was observed after 30 min at 25°C when using 1.12 U/μl T4 DNA Ligase. At 0.373 U/μl T4 DNA Ligase showed about 70% substrate turnover.

Properties

Specifications
Unit Definition

0.1 units is defined as the amount of enzyme that is needed to convert 1 pmol (of 18 pmol) of nicked DNA substrate in 20 minutes at 25 °C in a 20 µl reaction volume in a buffer consisting of 62.5 mM Tris-HCl pH 7.5, 10 mM DTT, 1 mM ATP, 0.05 mg/ml BSA and 25 mM KCl.

One Weiss Unit is equivalent to approximately 500 ArcticZymes Units.

Size

56.9 kDa

Inactivation

T4 DNA Ligase can be completely inactivated by incubating at 70°C for 10 minutes.

Storage and Stability

The enzyme is stable at -20°C for > 1 year in the supplied storage buffer.

The enzyme tolerates at least four freeze-thaw cycles (-80°C) without loss of activity.

Quality Control

ArcticZymes is dedicated to the quality of our products. T4 DNA Ligase is manufactured at our ISO 13485 certified facility in Norway.

dsDNA endonuclease activity

10 000 U T4 DNA Ligase 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

10 000 U T4 DNA Ligase 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.a

Exonuclease activity

10 000 U T4 DNA Ligase was incubated with either 3H-dATP labelled ds or ssDNA (0.5 µg, 500 bp) for 4 hours at 37°C. Acid soluble radioactivity from labelled DNA was not significantly over blank test for either substrate.

RNase activity

5000 U T4 DNA Ligase was incubated with a 2 kb RNA transcript (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of RNA degradation.

E. coli gDNA contamination

5000 U T4 DNA Ligase was analysed in a probe-based qPCR assay detecting the 23S ribosomal subunit in E. coli. No E. coli gDNA could be detected (LOD: < 3 E. coli genomic copies).

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
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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
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
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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
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
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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
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
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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
Request Quote
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
Request Quote
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
Request Quote
5
M-SAN HQ
70950-160
5 MU
>250 U/µl
$ 23,008.00 USD
22049.00
23008.00
Request Quote
5
dsDNase Triton Free
70601-110
100 kU
2 U/µl
$ 25,604.00 USD
22260.00
25604.00
Request Quote
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
Request Quote
6
SAN High Quality Triton Free
70921-150
500 kU
>250 U/µl
$ 5,767.00 USD
5011.00
5767.00
Request Quote
6
M-SAN HQ
70950-202
25 kU
25-30 U/µl
$ 606.00 USD
580.00
606.00
Request Quote
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
9
HL-dsDNase
70800-150
50 kU
50 U/µl
$ 30,330.00 USD
26368.00
30330.00
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
10
HL-dsDNase
70800-110
100 kU
2 U/µl
$ 45,552.00 USD
39608.00
45552.00
Request Quote
11
dsDNase
70600-110
100 kU
2 U/µl
$ 25,604.00 USD
22260.00
25604.00
Request Quote
11
Cod UNG
70500-151
50 kU
1 U/µl
$ 14,128.00 USD
12281.00
14128.00
Request Quote
11
HL-dsDNase
70800-120
500 kU
2 U/µl
$ 170,682.00 USD
148412.00
170682.00
Request Quote
12
Cod UNG
70500-150
50 kU
50 U/µl
$ 14,128.00 USD
12281.00
14128.00
Request Quote
12
dsDNase Glycerol Free
70610-50
5000 U
80 - 140 U/µl
$ 4,031.00 USD
3505.00
4031.00
Request Quote
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
Request Quote
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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T4 DNA Ligase
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