A novel approach for high-level expression and purification of GST-fused highly thermostable Taq DNA polymerase in Escherichia coli.

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.
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.

DNA polymerase ? (Rat)

10-102 100ug
EUR 923
Description: The DNA polymerase ? (Rat) is available in Europe and for worldwide shipping via Gentaur.

BIOTAQ DNA Polymerase

BIO-21040 500 Units Ask for price

IMMOLASE DNA Polymerase

BIO-21046 250 Units Ask for price

IMMOLASE DNA Polymerase

BIO-21047 500 Units Ask for price

IMMOLASE DNA Polymerase

BIO-21048 5000 Units Ask for price

ACCUZYME DNA Polymerase

BIO-21051 250 Units Ask for price

ACCUZYME DNA Polymerase

BIO-21052 500 Units Ask for price

BIOTAQ DNA Polymerase

BIO-21060 2500 Units Ask for price

MangoTaq DNA Polymerase

BIO-21078 5000 Units Ask for price

MangoTaq DNA Polymerase

BIO-21082 2000 Units Ask for price

MangoTaq DNA Polymerase

BIO-21083 1000 Units Ask for price

VELOCITY DNA Polymerase

BIO-21098 250 Units Ask for price

VELOCITY DNA Polymerase

BIO-21099 500 Units Ask for price

MyTaq DNA Polymerase

BIO-21105 500 units Ask for price

MyTaq DNA Polymerase

BIO-21105/S Sample Ask for price

MyTaq DNA Polymerase

BIO-21106 2500 units Ask for price

MyTaq DNA Polymerase

BIO-21107 5000 units Ask for price

MyFi DNA Polymerase

BIO-21117 250 Units Ask for price

MyFi DNA Polymerase

BIO-21117/S Sample Ask for price

MyFi DNA Polymerase

BIO-21118 500 Units Ask for price

MyFi DNA Polymerase

BIO-21119 2500 Units Ask for price

Ranger DNA Polymerase

BIO-21121 250 Units Ask for price

Ranger DNA Polymerase

BIO-21121/S Sample Ask for price

Ranger DNA Polymerase

BIO-21122 500 Units Ask for price

Taq DNA Polymerase

BA00104 1000U
EUR 123
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA Polymerase

BA00105 2500U
EUR 232
Description: High quality Taq polymerase for different PCR variations and downstream applications.

HotTaq DNA Polymerase

BA00203 500U
EUR 145
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq DNA Polymerase

BA00204 1000U
EUR 239
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq DNA Polymerase

BA00205 2500U
EUR 522
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

RedTaq DNA Polymerase

BA00303 500U
EUR 123
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.

RedTaq DNA Polymerase

BA00304 1000U
EUR 196
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.

RedTaq DNA Polymerase

BA00305 2500U
EUR 413
Description: High quality RedTaq polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00503 500U
EUR 145
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00504 1000U
EUR 239
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00505 2500U
EUR 522
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

Taq DNA polymerase

BT10101 500U
EUR 80
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA polymerase

BT10102 1000U
EUR 109
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA polymerase

BT10103 2000U
EUR 167
Description: High quality Taq polymerase for different PCR variations and downstream applications.

HotTaq DNA polymerase

BT10201 500U
EUR 120
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

HotTaq DNA polymerase

BT10202 1000U
EUR 190
Description: High quality HotTaq polymerase for different PCR variations and downstream applications.

Kodaq DNA Polymerase

G498 250 U (50 ul)
EUR 87

Kodaq DNA Polymerase

G499 1000 U (200 ul)
EUR 158

Taq DNA Polymerase

L7051001 1000 unit
EUR 195
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Taq DNA Polymerase

L7051200 200 unit
EUR 87
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Ready? DNA Polymerase

M1146-1000
EUR 332

Ready? DNA Polymerase

M1146-10000
EUR 1066

Ready? DNA Polymerase

M1146-5000
EUR 800

Breeze? DNA Polymerase

M1148-1000
EUR 1039

Breeze? DNA Polymerase

M1148-250
EUR 387

Distant? DNA Polymerase

M1150-1000
EUR 686

Distant? DNA Polymerase

M1150-250
EUR 278

Advance? DNA Polymerase

M1151-1000
EUR 746

Advance? DNA Polymerase

M1151-250
EUR 311

Outstretched? DNA Polymerase

M1152-1000
EUR 800

Outstretched? DNA Polymerase

M1152-250
EUR 332

T4 DNA Polymerase

M1211-100
EUR 321

Phi29 DNA Polymerase

M1239-1000
EUR 311

T4 DNA Polymerase

N101-01 2000 U
EUR 232

Taq DNA Polymerase

9001-2500
EUR 512

Taq DNA Polymerase

9001-500
EUR 191

PFU DNA Polymerase

9003-2500
EUR 512

PFU DNA Polymerase

9003-500
EUR 180

Laq? DNA Polymerase

9004-2500
EUR 610

Laq? DNA Polymerase

9004-500
EUR 218

AceTaq DNA Polymerase

P401-d1 250 U
EUR 136

AceTaq DNA Polymerase

P401-d2 1000 U
EUR 227

AceTaq DNA Polymerase

P401-d3 3000 U
EUR 447

100 BP DNA LADDER, 500UL PER KIT

M-DNA-100BP 1/pk
EUR 73
Description: Bioscience Mol Bio; DNA Ladder

1 KB DNA LADDER, 500UL PER KIT

M-DNA-1KB 1/pk
EUR 70
Description: Bioscience Mol Bio; DNA Ladder

Maximo Taq DNA Polymerase

S101 500 units
EUR 46

Maximo Taq DNA Polymerase

S102 5x500 units
EUR 107

Maximo Taq DNA Polymerase

S103 20x500 units
EUR 328

Taq DNA polymerase (+dNTPs)

02-001 200U
EUR 153
Description: The Taq DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Taq DNA polymerase (+dNTPs)

02-001-5 5×200U
EUR 257
Description: The Taq DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Taq DNA polymerase (-dNTPs)

02-011 200U
EUR 140
Description: The Taq DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Taq DNA polymerase (-dNTPs)

02-011-5 5×200U
EUR 218
Description: The Taq DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Pfu DNA polymerase (+dNTPs)

02-021 200U
EUR 179
Description: The Pfu DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Pfu DNA polymerase (+dNTPs)

02-021-5 5×200U
EUR 335
Description: The Pfu DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Pfu DNA polymerase (-dNTPs)

02-031 200U
EUR 166
Description: The Pfu DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Pfu DNA polymerase (-dNTPs)

02-031-5 5×200U
EUR 283
Description: The Pfu DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

DNA polymerase ? (C-His)

10-105 50ug
EUR 531
Description: The DNA polymerase ? (C-His) is available in Europe and for worldwide shipping via Gentaur.

HiFi DNA Polymerase wiodNTP

FYT033-500U 500U Ask for price

Accu DNA Polymerase wiodNTPs

FYT044-500U 500U Ask for price

amfiSure CloneEasy DNA Polymerase

P0325-010 100 units
EUR 232

amfiSure CloneEasy DNA Polymerase

P0325-025 250 units
EUR 401

MyTaq Red DNA Polymerase

BIO-21108 500 units Ask for price

MyTaq Red DNA Polymerase

BIO-21108/S Sample Ask for price

MyTaq Red DNA Polymerase

BIO-21109 2500 units Ask for price

MyTaq Red DNA Polymerase

BIO-21110 5000 units Ask for price

MyTaq HS DNA Polymerase

BIO-21111 250 units Ask for price

MyTaq HS DNA Polymerase

BIO-21111/S Sample Ask for price

MyTaq HS DNA Polymerase

BIO-21112 1000 units Ask for price

MyTaq HS DNA Polymerase

BIO-21113 2500 units Ask for price

DNA Polymerase gamma antibody

22870-100ul 100ul
EUR 390

DNA Polymerase beta Antibody

35404-100ul 100ul
EUR 390

DNA Polymerase β Antibody

33357-100ul 100ul
EUR 252

DNA Polymerase β Antibody

33357-50ul 50ul
EUR 187

DNA Polymerase ζ Antibody

33602-100ul 100ul
EUR 252

DNA Polymerase ζ Antibody

33602-50ul 50ul
EUR 187

DNA Polymerase θ Antibody

33611-100ul 100ul
EUR 252

DNA Polymerase θ Antibody

33611-50ul 50ul
EUR 187

DNA Polymerase α Antibody

33665-100ul 100ul
EUR 252

DNA Polymerase α Antibody

33665-50ul 50ul
EUR 187

DNA Polymerase λ Antibody

34098-100ul 100ul
EUR 252

DNA Polymerase λ Antibody

34098-50ul 50ul
EUR 187

DNA Polymerase beta Antibody

49605-100ul 100ul
EUR 333

DNA Polymerase beta Antibody

49605-50ul 50ul
EUR 239

Taq DNA Polymerase (500U)

9K-001-0001 500U
EUR 174.41

Taq DNA Polymerase (3000U)

9K-001-0002 3000U
EUR 485

Taq DNA Polymerase (6000U)

9K-001-0033 6000U
EUR 1028.75

Taq DNA Polymerase (1000U)

9K-001-0035 1000U
EUR 348.85

T DNA Polymerase Enzyme

abx071008-250U 250 U
EUR 189

T DNA Polymerase Enzyme

abx071008-3kU 3 kU
EUR 453

T DNA Polymerase Enzyme

abx071008-500U 500 U
EUR 217

TopTaq DNA Polymerase Enzyme

abx071014-250U 250 U
EUR 230

TopTaq DNA Polymerase Enzyme

abx071014-3kU 3 kU
EUR 787

TopTaq DNA Polymerase Enzyme

abx071014-500U 500 U
EUR 300

EasyPfu DNA Polymerase Enzyme

abx071016-250U 250 U
EUR 189

EasyPfu DNA Polymerase Enzyme

abx071016-3kU 3 kU
EUR 439

EasyPfu DNA Polymerase Enzyme

abx071016-500U 500 U
EUR 217

FastPfu DNA Polymerase Enzyme

abx071018-250U 250 U
EUR 258

FastPfu DNA Polymerase Enzyme

abx071018-3kU 3 kU
EUR 926

FastPfu DNA Polymerase Enzyme

abx071018-500U 500 U
EUR 314

Taq DNA Polymerase (Recombinant)

abx073521-10000U 10000 U
EUR 592

Taq DNA Polymerase (Recombinant)

abx073521-1000U 1000 U
EUR 230

Taq DNA Polymerase (Recombinant)

abx073521-3000U 3000 U
EUR 328

Pfu DNA Polymerase (Recombinant)

abx074017-100U 100 U
EUR 328

Pfu DNA Polymerase (Recombinant)

abx074017-2500U 2500 U
EUR 1274

Pfu DNA Polymerase (Recombinant)

abx074017-500U 500 U
EUR 439

DNA Polymerase beta Antibody

20-abx008852
  • EUR 300.00
  • EUR 439.00
  • EUR 189.00
  • 100 ul
  • 200 ul
  • 30 ul

DNA Polymerase beta Antibody

20-abx013060
  • EUR 314.00
  • EUR 98.00
  • EUR 398.00
  • EUR 495.00
  • 100 ug
  • 10 ug
  • 200 ug
  • 300 µg

DNA Polymerase alpha Antibody

20-abx013375
  • EUR 314.00
  • EUR 98.00
  • EUR 398.00
  • EUR 495.00
  • 100 ug
  • 10 ug
  • 200 ug
  • 300 µg

Recombinant Pfu DNA Polymerase

7-03322 100U Ask for price

Recombinant Pfu DNA Polymerase

7-03323 500U Ask for price

Recombinant Pfu DNA Polymerase

7-03324 2500U Ask for price

Recombinant Taq DNA Polymerase

7-03634 1,000U Ask for price

Recombinant Taq DNA Polymerase

7-03635 3,000U Ask for price

Recombinant Taq DNA Polymerase

7-03636 10,000U Ask for price

DNA polymerase beta Antibody

abx232432-100ug 100 ug
EUR 481

Taq DNA Polymerase, 5units/㎕

T2200-050 500units
EUR 131

Taq DNA Polymerase, 5units/㎕

T2200-100 2x500 units
EUR 192

Taq DNA Polymerase, 5units/㎕

T2200-200 4x500 units
EUR 313

DNA Polymerase alpha Antibody

AF5235 200ul
EUR 304
Description: DNA Polymerase α Antibody detects endogenous levels of total DNA Polymerase α.

DNA Polymerase beta Antibody

ABF0148 100 ug
EUR 438

Taq DNA Polymerase (PCR)

G008 5000 U (1.0 ml)
EUR 205

Taq DNA Polymerase (PCR)

G009 1000 U (200 ul)
EUR 87

HotStart DNA Polymerase (PCR)

G011 250 U (50 ul)
EUR 82

HotStart DNA Polymerase (PCR)

G039 1000 U (200 ul)
EUR 158

Taq DNA Polymerase (PCR)

G126 10000 U (2 x 1.0 ml)
EUR 309

TaqFast DNA Polymerase (PCR)

G277 250 U (50 ul)
EUR 73

TaqFast DNA Polymerase (PCR)

G278 1000 U (200 ul)
EUR 134

Bloodirect DNA Polymerase (PCR)

G462 100 U (100 ul)
EUR 61

Bloodirect DNA Polymerase (PCR)

G463 400 U (400 ul)
EUR 87

SensTaq HotStart DNA Polymerase

G937 250U
EUR 87

SensTaq HotStart DNA Polymerase

G938 1000U
EUR 158

DNA Polymerase alpha Antibody

DF3082 200ul
EUR 304
Description: DNA Polymerase α Antibody detects endogenous levels of total DNA Polymerase α.

DNA Polymerase beta Antibody

AF0148 200ul
EUR 304
Description: DNA Polymerase beta antibody detects endogenous levels of total DNA Polymerase beta.

DNA Polymerase beta antibody

70R-31161 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase beta antibody

DNA Polymerase alpha antibody

70R-31578 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase alpha antibody

DNA Polymerase lambda antibody

70R-32186 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase lambda antibody

DNA Polymerase zeta antibody

70R-33711 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase zeta antibody

DNA Polymerase zeta antibody

70R-33727 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase zeta antibody

DNA Polymerase theta antibody

70R-33737 100 ug
EUR 327
Description: Rabbit polyclonal DNA Polymerase theta antibody

DNA polymerase gamma antibody

70R-12532 100 ul
EUR 457
Description: Affinity purified Rabbit polyclonal DNA polymerase gamma antibody

DNA Polymerase beta antibody

70R-50226 100 ul
EUR 244
Description: Purified Polyclonal DNA Polymerase beta antibody

Hotstart Taq DNA Polymerase

L7052001 1000 units
EUR 568
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

Hotstart Taq DNA Polymerase

L7052250 250 units
EUR 195
Description: Premade ready to use kits will always come in handy. Get your experiment done right form the first try by using a validated kit with perfectly balanced reagents proportions and compatibility and by following a clear protocol.

anti-DNA polymerase beta

YF-PA13864 50 ug
EUR 363
Description: Mouse polyclonal to DNA polymerase beta

anti-DNA polymerase beta

YF-PA13865 100 ul
EUR 403
Description: Rabbit polyclonal to DNA polymerase beta

anti-DNA polymerase eta

YF-PA13871 50 ug
EUR 363
Description: Mouse polyclonal to DNA polymerase eta

anti-DNA polymerase eta

YF-PA13872 100 ug
EUR 403
Description: Rabbit polyclonal to DNA polymerase eta

Robust Ready? DNA Polymerase

M1147-1000
EUR 958

Robust Ready? DNA Polymerase

M1147-250
EUR 365

Fire Start? DNA Polymerase

M1149-1000
EUR 637

Fire Start? DNA Polymerase

M1149-250
EUR 256

Blood Advance? DNA Polymerase

M1153-100
EUR 224

Blood Advance? DNA Polymerase

M1153-400
EUR 474

anti-DNA polymerase alpha

YF-PA24412 50 ul
EUR 334
Description: Mouse polyclonal to DNA polymerase alpha


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.