Polymerases are enzymes that synthesize long chains or polymers of nucleic acids, including DNA or RNA of nucleotides. They assemble to copy the template nucleic acids DNA or RNA strand using the base-pairing interactions. One of the polymerase enzyme, Taq DNA polymerase, originally isolated from Thermus aquaticus (Taq) is an enzyme widely used in molecular biology so far.
The properties of thermostable enzymes has contributed majorly to specificity, automation, and efficacy of the polymerase chain reaction (PCR), making it a powerful tool for molecular biology research today throughout the world. Purification of Taq DNA polymerase from the original host results in low yields, labor and time consumption.
Therefore, much research has been done before to get this enzyme using an alternative host. So far, all of the existing methodology is more laborious, time consuming and requires a heavy cost. We use a new approach to purifying the enzyme with relatively high efficiency, yield and minimum time consumption using Escherichia coli (E. coli) as an alternative host.
We cloned 2500 base pairs of DNA Taq polymerase gene into pGEX-4T-1 vector containing GST-tag, downstream of tac promoter and expressed using isopropyl β-D-1-thiogalactopyranoside (IPTG) as an inducer. The enzyme was purified using chromatography efficient new approach and use in routine PCR tests in our laboratory. Our findings suggest a new approach to facilitate the availability of polymerases for molecular and diagnostic studies. In the future, may be used for purification of other recombinant peptides or proteins used in structural biology and proteomics research-based.
A novel approach for high-level expression and purification of GST-fused highly thermostable Taq DNA polymerase in Escherichia coli.
Outside Chelation: EDTA binds Closely Taq DNA Polymerase, Mutt and dUTPase and direct Inhibiting Activity dNTPase.
EDTA is commonly used as a metal ion chelator efficient enzyme cofactor. It is highly soluble, optically inactive and do not interfere with most chemicals used in making standard buffer EDTA common options for generating metal-free conditions for biochemical and biophysical investigations.
However, controversy in the literature about the activity of the metal-free enzyme is achieved by using EDTA or otherwise called our attention to the alleged effects of external EDTA chelation. Here, we show that EDTA compete for sites of hydrolases dUTPase nucleotide nucleotide binding by developing a network of interactions within the active site similar to the substrate.
To reach these findings, we apply the kinetics and molecular docking technique uses two different dUTPases. Furthermore, we directly measured the binding of EDTA to dUTPases and two other dNTPases, Taq polymerase and Mutt using isothermal titration calorimetry. EDTA binds proven exothermic and enthalpy driven primarily by submicromolar dissociation constant is much lower than with an enzyme: substrate or Mg: EDTA complex. Control of protein, including ATPase, do not interact with EDTA. Our findings indicate that EDTA can act as a selective inhibitor of the enzyme hydrolysis of dNTPs and urges a rethinking of the use of EDTA in enzymatic trial.
Taq DNA Polymerase
BA00105
Bioatlas
2500U
EUR 278.4
Description: High quality Taq polymerase for different PCR variations and downstream applications.
Pfu DNA Polymerase
BA00503
Bioatlas
500U
EUR 174
Description: High quality Pfu polymerase for different PCR variations and downstream applications.
Pfu DNA Polymerase
BA00504
Bioatlas
1000U
EUR 286.8
Description: High quality Pfu polymerase for different PCR variations and downstream applications.
Pfu DNA Polymerase
BA00505
Bioatlas
2500U
EUR 626.4
Description: High quality Pfu polymerase for different PCR variations and downstream applications.
Taq DNA polymerase
BT10101
Bioatlas
500U
EUR 96
Description: High quality Taq polymerase for different PCR variations and downstream applications.
Taq DNA polymerase
BT10102
Bioatlas
1000U
EUR 130.8
Description: High quality Taq polymerase for different PCR variations and downstream applications.
Taq DNA polymerase
BT10103
Bioatlas
2000U
EUR 200.4
Description: High quality Taq polymerase for different PCR variations and downstream applications.
Taq DNA Polymerase
9001-2500
Biovision
each
EUR 614.4
Taq DNA Polymerase
9001-500
Biovision
each
EUR 229.2
PFU DNA Polymerase
9003-2500
Biovision
each
EUR 614.4
PFU DNA Polymerase
9003-500
Biovision
each
EUR 216
Laq? DNA Polymerase
9004-2500
Biovision
each
EUR 732
Laq? DNA Polymerase
9004-500
Biovision
each
EUR 261.6
Taq DNA Polymerase
L7051001
Biochain
1000 unit
EUR 159
Taq DNA Polymerase
L7051200
Biochain
200 unit
EUR 41
Pfu DNA Polymerase
S116
GeneOn
1x250 units
EUR 91.2
Pfu DNA Polymerase
S117
GeneOn
2x250 units
EUR 112.8
Pfu DNA Polymerase
S118
GeneOn
10x250 units
EUR 363.6
Bst DNA Polymerase
S600
GeneOn
2000 U
EUR 128.4
MyFi DNA Polymerase
BIO-21117
Bioline
250 Units
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MyFi DNA Polymerase
BIO-21117/S
Bioline
Sample
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MyFi DNA Polymerase
BIO-21118
Bioline
500 Units
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MyFi DNA Polymerase
BIO-21119
Bioline
2500 Units
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Taq DNA Polymerase (3000U)
9K-001-0002
Bio Basic
3000U
EUR 582
Taq DNA Polymerase (6000U)
9K-001-0033
Bio Basic
6000U
EUR 1234.5
Taq DNA Polymerase (1000U)
9K-001-0035
Bio Basic
1000U
EUR 418.62
MyTaq DNA Polymerase
BIO-21105
Bioline
500 units
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MyTaq DNA Polymerase
BIO-21105/S
Bioline
Sample
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MyTaq DNA Polymerase
BIO-21106
Bioline
2500 units
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MyTaq DNA Polymerase
BIO-21107
Bioline
5000 units
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Ready? DNA Polymerase
M1146-1000
Biovision
each
EUR 398.4
Ready? DNA Polymerase
M1146-10000
Biovision
each
EUR 1279.2
Ready? DNA Polymerase
M1146-5000
Biovision
each
EUR 960
Phi29 DNA Polymerase
M1239-1000
Biovision
each
EUR 373.2
Kodaq DNA Polymerase
G498
ABM
250 U (50 ul)
EUR 104.4
Kodaq DNA Polymerase
G499
ABM
1000 U (200 ul)
EUR 189.6
HotTaq DNA Polymerase
BA00203
Bioatlas
500U
EUR 174
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.
HotTaq DNA Polymerase
BA00204
Bioatlas
1000U
EUR 286.8
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.
HotTaq DNA Polymerase
BA00205
Bioatlas
2500U
EUR 626.4
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.
RedTaq DNA Polymerase
BA00303
Bioatlas
500U
EUR 147.6
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.
RedTaq DNA Polymerase
BA00304
Bioatlas
1000U
EUR 235.2
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.
RedTaq DNA Polymerase
BA00305
Bioatlas
2500U
EUR 495.6
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.
BIOTAQ DNA Polymerase
BIO-21040
Bioline
500 Units
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BIOTAQ DNA Polymerase
BIO-21060
Bioline
2500 Units
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Ranger DNA Polymerase
BIO-21121
Bioline
250 Units
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Ranger DNA Polymerase
BIO-21121/S
Bioline
Sample
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Ranger DNA Polymerase
BIO-21122
Bioline
500 Units
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HotTaq DNA polymerase
BT10201
Bioatlas
500U
EUR 144
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.
HotTaq DNA polymerase
BT10202
Bioatlas
1000U
EUR 228
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.
AceTaq DNA Polymerase
P401-d1
Vazyme
250 U
EUR 163.2
AceTaq DNA Polymerase
P401-d2
Vazyme
1000 U
EUR 272.4
AceTaq DNA Polymerase
P401-d3
Vazyme
3000 U
EUR 536.4
Breeze? DNA Polymerase
M1148-1000
Biovision
each
EUR 1246.8
Breeze? DNA Polymerase
M1148-250
Biovision
each
EUR 464.4
AceTaq DNA Polymerase
P401-MD1
Vazyme
250 U
EUR 46.4
Taq DNA Polymerase (500U)
9K-001-0001
Bio Basic
500U
EUR 209.29
Distant? DNA Polymerase
M1150-1000
Biovision
each
EUR 823.2
Distant? DNA Polymerase
M1150-250
Biovision
each
EUR 333.6
Advance? DNA Polymerase
M1151-1000
Biovision
each
EUR 895.2
Advance? DNA Polymerase
M1151-250
Biovision
each
EUR 373.2
IMMOLASE DNA Polymerase
BIO-21046
Bioline
250 Units
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IMMOLASE DNA Polymerase
BIO-21047
Bioline
500 Units
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IMMOLASE DNA Polymerase
BIO-21048
Bioline
5000 Units
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ACCUZYME DNA Polymerase
BIO-21051
Bioline
250 Units
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ACCUZYME DNA Polymerase
BIO-21052
Bioline
500 Units
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MangoTaq DNA Polymerase
BIO-21078
Bioline
5000 Units
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MangoTaq DNA Polymerase
BIO-21082
Bioline
2000 Units
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MangoTaq DNA Polymerase
BIO-21083
Bioline
1000 Units
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VELOCITY DNA Polymerase
BIO-21098
Bioline
250 Units
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VELOCITY DNA Polymerase
BIO-21099
Bioline
500 Units
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Taq HS DNA Polymerase
P132-d1
Vazyme
500 U
EUR 38.28
Taq HS DNA Polymerase
P132-d2
Vazyme
1,000 U
EUR 71.34
Taq HS DNA Polymerase
P132-d3
Vazyme
5,000 U
EUR 324.8
Outstretched? DNA Polymerase
M1152-1000
Biovision
each
EUR 960
Outstretched? DNA Polymerase
M1152-250
Biovision
each
EUR 398.4
DNA polymerase ? (C-His)
10-105
Sceti
50ug
EUR 637.2
Description: The DNA polymerase ? (C-His) is available in Europe and for worldwide shipping via Gentaur.
Taq DNA Polymerase (PCR)
G008
ABM
5000 U (1.0 ml)
EUR 246
Taq DNA Polymerase (PCR)
G009
ABM
1000 U (200 ul)
EUR 104.4
Taq DNA Polymerase (PCR)
G126
ABM
10000 U (2 x 1.0 ml)
EUR 370.8
MyTaq HS DNA Polymerase
BIO-21111
Bioline
250 units
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MyTaq HS DNA Polymerase
BIO-21111/S
Bioline
Sample
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MyTaq HS DNA Polymerase
BIO-21112
Bioline
1000 units
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MyTaq HS DNA Polymerase
BIO-21113
Bioline
2500 units
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Taq Plus DNA Polymerase
P201-01
Vazyme
250 U
EUR 147.6
Taq Plus DNA Polymerase
P201-02
Vazyme
1000 U
EUR 213.6
Taq Plus DNA Polymerase
P201-03
Vazyme
3000 U
EUR 382.8
ACTaq? Taq DNA Polymerase, 1000U
E2100-1000U
ACTGene
each
EUR 260.4
Taq DNA polymerase (+dNTPs)
02-001
Sceti
200U
EUR 183.6
Description: The Taq DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.
Taq DNA polymerase (+dNTPs)
02-001-5
Sceti
5×200U
EUR 308.4
Description: The Taq DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.
Taq DNA polymerase (-dNTPs)
02-011
Sceti
200U
EUR 168
Description: The Taq DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.
Taq DNA polymerase (-dNTPs)
02-011-5
Sceti
5×200U
EUR 261.6
Description: The Taq DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.
Pfu DNA polymerase (+dNTPs)
02-021
Sceti
200U
EUR 214.8
Description: The Pfu DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.
Pfu DNA polymerase (+dNTPs)
02-021-5
Sceti
5×200U
EUR 402
Description: The Pfu DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.
Pfu DNA polymerase (-dNTPs)
02-031
Sceti
200U
EUR 199.2
Description: The Pfu DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.
Pfu DNA polymerase (-dNTPs)
02-031-5
Sceti
5×200U
EUR 339.6
Description: The Pfu DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.
T DNA Polymerase Enzyme
abx071008-250U
Abbexa
250 U
EUR 226.8
T DNA Polymerase Enzyme
abx071008-3kU
Abbexa
3 kU
EUR 543.6
T DNA Polymerase Enzyme
abx071008-500U
Abbexa
500 U
EUR 260.4
MyTaq Red DNA Polymerase
BIO-21108
Bioline
500 units
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MyTaq Red DNA Polymerase
BIO-21108/S
Bioline
Sample
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MyTaq Red DNA Polymerase
BIO-21109
Bioline
2500 units
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MyTaq Red DNA Polymerase
BIO-21110
Bioline
5000 units
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ACTaq? Long DNA Polymerase, 1000U
E2200-1000U
ACTGene
each
EUR 282
Fire Start? DNA Polymerase
M1149-1000
Biovision
each
EUR 764.4
Fire Start? DNA Polymerase
M1149-250
Biovision
each
EUR 307.2
Maximo Taq DNA Polymerase
S101
GeneOn
500 units
EUR 55.2
Maximo Taq DNA Polymerase
S102
GeneOn
5x500 units
EUR 128.4
Maximo Taq DNA Polymerase
S103
GeneOn
20x500 units
EUR 393.6
Taq DNA Polymerase, 5units/㎕
T2200-050
GenDepot
500units
EUR 157.2
Taq DNA Polymerase, 5units/㎕
T2200-100
GenDepot
2x500 units
EUR 230.4
Taq DNA Polymerase, 5units/㎕
T2200-200
GenDepot
4x500 units
EUR 375.6
ACTaq? Taq DNA Polymerase, 250U
E2100-250U
ACTGene
each
EUR 122.4
DNA Polymerase λ Antibody
34098-100ul
SAB
100ul
EUR 302.4
DNA Polymerase λ Antibody
34098-50ul
SAB
50ul
EUR 224.4
DNA Polymerase β Antibody
33357-100ul
SAB
100ul
EUR 302.4
DNA Polymerase β Antibody
33357-50ul
SAB
50ul
EUR 224.4
DNA Polymerase ζ Antibody
33602-100ul
SAB
100ul
EUR 302.4
DNA Polymerase ζ Antibody
33602-50ul
SAB
50ul
EUR 224.4
DNA Polymerase θ Antibody
33611-100ul
SAB
100ul
EUR 302.4
DNA Polymerase θ Antibody
33611-50ul
SAB
50ul
EUR 224.4
DNA Polymerase α Antibody
33665-100ul
SAB
100ul
EUR 302.4
DNA Polymerase α Antibody
33665-50ul
SAB
50ul
EUR 224.4
TaqFast DNA Polymerase (PCR)
G277
ABM
250 U (50 ul)
EUR 87.6
TaqFast DNA Polymerase (PCR)
G278
ABM
1000 U (200 ul)
EUR 160.8
Fast-Taq DNA Polymerase (500U)
9K-001-0036
Bio Basic
500U
EUR 420.65
ACTaq? Long DNA Polymerase, 250U
E2200-250U
ACTGene
each
EUR 127.2
Hotstart Taq DNA Polymerase
L7052001
Biochain
1000 units
EUR 567
Hotstart Taq DNA Polymerase
L7052250
Biochain
250 units
EUR 159
Robust Ready? DNA Polymerase
M1147-1000
Biovision
each
EUR 1149.6
Robust Ready? DNA Polymerase
M1147-250
Biovision
each
EUR 438
HotStart DNA Polymerase (PCR)
G011
ABM
250 U (50 ul)
EUR 98.4
HotStart DNA Polymerase (PCR)
G039
ABM
1000 U (200 ul)
EUR 189.6
T4 DNA Polymerase, 5u/ul
BEP0061
Bio Basic
100U
EUR 153.97
T4 DNA Polymerase (5,000 U/ml)
RK20539
Abclonal
25 U
Ask for price
DNA polymerase beta Antibody
abx232432-100ug
Abbexa
100 ug
EUR 577.2
TopTaq DNA Polymerase Enzyme
abx071014-250U
Abbexa
250 U
EUR 276
TopTaq DNA Polymerase Enzyme
abx071014-3kU
Abbexa
3 kU
EUR 944.4
TopTaq DNA Polymerase Enzyme
abx071014-500U
Abbexa
500 U
EUR 360
DNA Polymerase beta Antibody
AF0148
Affbiotech
200ul
EUR 420
DNA Polymerase beta Antibody
49605-100ul
SAB
100ul
EUR 399.6
DNA Polymerase beta Antibody
49605-50ul
SAB
50ul
EUR 286.8
DNA Polymerase beta antibody
70R-31161
Fitzgerald
100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase beta antibody
DNA Polymerase zeta antibody
70R-33711
Fitzgerald
100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase zeta antibody
DNA Polymerase zeta antibody
70R-33727
Fitzgerald
100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase zeta antibody
DNA Polymerase beta antibody
70R-50226
Fitzgerald
100 ul
EUR 292.8
Description: Purified Polyclonal DNA Polymerase beta antibody
DNA Polymerase beta Antibody
35404-100ul
SAB
100ul
EUR 468
DNA Polymerase beta Antibody
20-abx008852
Abbexa
EUR 360.00
EUR 526.80
EUR 226.80
DNA Polymerase beta Antibody
20-abx013060
Abbexa
EUR 376.80
EUR 117.60
EUR 477.60
EUR 594.00
100 ug
10 ug
200 ug
300 µg
DNA Polymerase beta Antibody
DF6552
Affbiotech
200ul
EUR 420
Champagne Taq DNA Polymerase
P122-d1
Vazyme
500 U (2.5 U/μl)
EUR 212.4
Champagne Taq DNA Polymerase
P122-d2
Vazyme
500 U (5 U/μl)
EUR 212.4
Champagne Taq DNA Polymerase
P122-d3
Vazyme
500 U (10 U/μl)
EUR 212.4
Blood Advance? DNA Polymerase
M1153-100
Biovision
each
EUR 268.8
Blood Advance? DNA Polymerase
M1153-400
Biovision
each
EUR 568.8
Taq DNA Polymerase, 5u/ul
B0089(D0089)
Bio Basic
5X200U
EUR 112.2
Pfu DNA Polymerase, 5u/ul
B0093(D0090P)
Bio Basic
500U
EUR 164.4
DNA Polymerase I, 10u/ul
BEP0041
Bio Basic
500U
EUR 128.9
Tsg DNA Polymerase, 5u/ul
D0081
Bio Basic
5X200U
EUR 112.2
DNA Polymerase gamma antibody
22870-100ul
SAB
100ul
EUR 468
EasyPfu DNA Polymerase Enzyme
abx071016-250U
Abbexa
250 U
EUR 226.8
EasyPfu DNA Polymerase Enzyme
abx071016-3kU
Abbexa
3 kU
EUR 526.8
EasyPfu DNA Polymerase Enzyme
abx071016-500U
Abbexa
500 U
EUR 260.4
FastPfu DNA Polymerase Enzyme
abx071018-250U
Abbexa
250 U
EUR 309.6
FastPfu DNA Polymerase Enzyme
abx071018-3kU
Abbexa
3 kU
EUR 1111.2
FastPfu DNA Polymerase Enzyme
abx071018-500U
Abbexa
500 U
EUR 376.8
DNA Polymerase alpha Antibody
AF5235
Affbiotech
200ul
EUR 420
DNA polymerase gamma antibody
70R-12532
Fitzgerald
100 ul
EUR 548.4
Description: Affinity purified Rabbit polyclonal DNA polymerase gamma antibody
DNA Polymerase alpha antibody
70R-31578
Fitzgerald
100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase alpha antibody
DNA Polymerase theta antibody
70R-33737
Fitzgerald
100 ug
EUR 392.4
Description: Rabbit polyclonal DNA Polymerase theta antibody
DNA Polymerase alpha Antibody
20-abx013375
Abbexa
EUR 376.80
EUR 117.60
EUR 477.60
EUR 594.00
100 ug
10 ug
200 ug
300 µg
DNA Polymerase alpha Antibody
DF3082
Affbiotech
200ul
EUR 420
DNA Polymerase theta Antibody
DF13563
Affbiotech
100ul
EUR 420
Bloodirect DNA Polymerase (PCR)
G462
ABM
100 U (100 ul)
EUR 73.2
In order to develop a test to identify the genes responsible for antibiotic resistance in gram-negative bacteria, it has been found that the standard of Taq DNA polymerase (not DNA-free) blaTEM contaminated with fragments of genes that varied in length and number. BlaTEM complete gene sequences are absent or detected in very small amounts. We developed an approach to avoid false positive findings caused by contamination in the polymerase gene sequence blaTEM conventional.