Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans-Expression and characterization.

Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans-Expression and characterization.

DNA polymerases are present in all organisms and enzymes important that synthesize DNA molecules. They are used in various fields of science, especially as an important component for the in vitro synthesis of DNA, known as PCR. modern diagnostics, molecular biology and genetic engineering of DNA polymerase needs which demonstrate improved performance. This study aims to get NeqSSB-TaqS new DNA polymerase fusion of domain Stoffel Taq DNA and single-stranded DNA binding protein from Nanoarchaeum equitans as to significantly improve the properties of the DNA polymerase.

Stoffel Taq DNA coding sequence specific DNA polymerase and DNA-binding proteins Nanoarchaeum equitans (NeqSSB-like protein) fused. A new recombinant genes obtained were cloned into pET-30 Ek / LIC vector and introduced into E. coli for expression. Purified recombinant enzymes and enzymatic properties including the activity of DNA polymerase, PCR amplification level, thermostability, processivity and resistance to inhibitors, were tested. The results of the target protein around 18 mg / l after 24 hours of IPTG induction.

The specific activity of the polymerase is 2200 U / mg. Recombinant NeqSSB-TaqS exhibited a higher level extensions (1000 bp template 20 s), processivity (19 nt), thermostability (half-life 35 minutes at 95 ° C) and a higher tolerance for PCR inhibitors (0.3 to 1.25 % of whole blood, 0.84 to 13.5 ug heparin lactoferrin and 4.7 to 150 ng) of Taq DNA polymerase Stoffel. In addition, our research shows that NeqSSB-TaqS DNA polymerase has a high degree of flexibility in relation to the Mg 2+ ions (from 1 to 5 mM) and KCl or (NH4) 2SO4 salt (more than 60 mM and 40 mM, respectively ). NeqSSB-TaqS using DNA polymerase Taq DNA polymerase can not be a better choice in many applications of PCR.

 Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans-Expression and characterization.
Fusion of Taq DNA polymerase with single-stranded DNA binding-like protein of Nanoarchaeum equitans-Expression and characterization.

dUTPs zwitterionic conjugated with Cy3 or Cy5 analog Fluorophore effective substrate for DNA amplification and labeling by Taq polymerase.

To develop the structural modifications dNTP compatible with Taq DNA polymerase activity, we synthesized eight dUTP derivatives conjugated with Cy3 or Cy5 dyes of different analog responsible and charge distribution throughout the fluorophore. Cy3- derivatives and commercial dUTP and Cy5-dUTP studied in Taq polymerase dependent polymerase chain reaction (PCR) and the primer extension reaction using a model template that contains one, two and three adjacent nucleotides adenine.

The relative amount of DNA is amplified and kinetic parameters Km and Vmax characterize grief labeled incorporation have been estimated using fluorescence measurements and analyzed. The dUTPs label with analog zwitterionic electroneutral of Cy3 or Cy5 fluorophores Taq polymerase is used by approximately one order of magnitude more effective than dUTPs analog labeled with a negatively charged of Cy3 or Cy5. Nucleotidyl transferase activity of Taq polymerase were also observed and resulted in the addition dumps electroneutral labeled with fluorophores or positively charged to the 3 ‘end of the DNA.

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Description: High quality Taq polymerase for different PCR variations and downstream applications.

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Description: High quality Taq polymerase for different PCR variations and downstream applications.

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Description: High quality Taq polymerase for different PCR variations and downstream applications.

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Description: High quality Taq polymerase for different PCR variations and downstream applications.

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Taq DNA polymerase (+dNTPs)

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Description: The Taq DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

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Description: The Taq DNA polymerase (+dNTPs) is available in Europe and for worldwide shipping via Gentaur.

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Description: The Taq DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

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Description: The Taq DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

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EUR 212.4

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Taq Plus DNA Polymerase

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GR108007 200 tests
EUR 230

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Taq Plus DNA Polymerase (PCR)

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EUR 87.6

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G040 1000 U (200 ul)
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ACTaq? Taq Blue DNA Polymerase, 1000U

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Taq DNA Polymerase (Mg2+ plus Buffer)

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EUR 314.4

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EUR 145.2

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EUR 231.6

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Cheetah Hot Start Taq DNA Polymerase

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EUR 355.2
Description: Minimum order quantity: 1 unit of 500EU

Taq Plus DNA Polymerase (Recombinant)

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EUR 71.2

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Taq DNA Polymerase with dNTP Mix (6000U)

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Taq DNA Polymerase (Mg2+ plus Buffer, with dNTP)

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EUR 163.2

Taq DNA Polymerase (Mg2+ plus Buffer, with dNTP)

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Taq DNA Polymerase (Mg2+ plus Buffer, with dNTP)

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Taq DNA Polymerase (Mg2+ free Buffer, with dNTP)

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EUR 163.2

Taq DNA Polymerase (Mg2+ free Buffer, with dNTP)

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Reversible 1-min Hot Start Taq DNA Polymerase

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Taq DNA Polymerase - 5,000 units with separate dNTP Mix

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EUR 802.8

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EUR 399.6

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EUR 286.8

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EUR 122.4

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Taq Pro Multiplex DNA Polymerase (High sensitivity)

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EUR 290

Maximo Blue-Taq DNA Polymerase (ready-to-load, conc. 1u/µl)

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EUR 63.6

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Hot Start Taq DNA Polymerase - 1000 units with seperate dNTP mix

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amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2

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EUR 200.4

amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2

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amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2

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Maximo Taq DNA Polymerase (2X pre-mix, ready-to-use)

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Maximo H-SPlus Taq DNA Polymerase (for Hot-Start PCR)

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EUR 86.4

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EUR 231.6

amfiEco Taq DNA Polymerase, 1 unit/ul,Including dNTP&MgCl2

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Anti-Taq DNA Polymerase Conjugated Antibody

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EUR 476.4

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9K-005-0020 500U
EUR 245.83

Accuris Taq Polymerase, 1000 units

PR1000-1000 1 PC
EUR 167.1

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PR1000-6000 1 PC
EUR 853.3

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PR1000-500 1 PC
EUR 103.6

TaqFast DNA Polymerase (PCR)

G277 250 U (50 ul)
EUR 87.6

TaqFast DNA Polymerase (PCR)

G278 1000 U (200 ul)
EUR 160.8

DNA Polymerase

ABD3082 100 ug
EUR 525.6

DNA Polymerase

ABF5235 100 ug
EUR 525.6

Fast-Taq DNA Polymerase with 10mM dNTP Mix (500U)

9K-001-0003 500U
EUR 575.22

Fast-Taq DNA Polymerase with 10mM dNTP Mix (2500U)

9K-001-0004 2500U
EUR 2465.38

High-Taq DNA Polymerase with 10mM dNTP Mix (250U)

9K-001-0005 250U
EUR 384.17

High-Taq DNA Polymerase with 10mM dNTP Mix (5000U)

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EUR 2878.8

High-Taq DNA Polymerase with 10mM dNTP Mix (4x250U)

9K-001-0006 4x250U
EUR 1055.98

Tsg PlusDNA Polymerase, 5u/ul

D0088 200U
EUR 94.8

T4 DNA Polymerase

N101-01 2000 U
EUR 278.4

T4 DNA Polymerase

M1211-100 each
EUR 385.2

DNA polymerase ? (Rat)

10-101 20ug
EUR 465.6
Description: The DNA polymerase ? (Rat) is available in Europe and for worldwide shipping via Gentaur.

DNA polymerase ? (Rat)

10-102 100ug
EUR 1107.6
Description: The DNA polymerase ? (Rat) is available in Europe and for worldwide shipping via Gentaur.

Pfu DNA Polymerase

BA00503 500U
EUR 174
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00504 1000U
EUR 286.8
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

Pfu DNA Polymerase

BA00505 2500U
EUR 626.4
Description: High quality Pfu polymerase for different PCR variations and downstream applications.

PFU DNA Polymerase

9003-2500 each
EUR 614.4

PFU DNA Polymerase

9003-500 each
EUR 216

Laq? DNA Polymerase

9004-2500 each
EUR 732

Laq? DNA Polymerase

9004-500 each
EUR 261.6

Pfu DNA Polymerase

S116 1x250 units
EUR 91.2

Pfu DNA Polymerase

S117 2x250 units
EUR 112.8

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S118 10x250 units
EUR 363.6

Bst DNA Polymerase

S600 2000 U
EUR 128.4

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

Accuris Taq Polymerase, sample, 50 units

PR1000-S 1 PC
EUR 7.1

MyTaq DNA Polymerase

BIO-21105 500 units Ask for price

The introduction of mutual compensation costs to fluorophores or other functional groups dNTP conjugation can be considered as a basis for the preparation of modified nucleoside PCR-compatible triphosphates.Using Sso7d from Sulfolobus solfataricus as DNA binding protein fusion Taq DNA polymerase at the amino end, we reported hyper-expression and purification methodology novel Sso7d-Taq polymerase (S-Taq) using a two-phase system of water extraction followed by Ni-affinity chromatography.

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