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Mutant Taq DNA polymerases with improved elongation ability as a useful reagent for genetic engineering.

Mutant Taq DNA polymerases with improved elongation ability as a useful reagent for genetic engineering.

DNA polymerase is widely used for manipulating DNA in vitro, including DNA cloning, sequencing, DNA labeling, mutagenesis, and other experiments. thermostable DNA polymerase is very useful and can be quite worthwhile after the development of PCR technology. A DNA polymerases from Thermus aquaticus (Taq polymerase) is the most well-known DNA polymerase as PCR enzyme, and has been widely used around the world. In this study, the gene fragment of family A DNA polymerase was amplified by PCR from the DNA of microorganisms in environmental soil samples, using a set of primers for two conserved regions.

The corresponding region of the pol gene for Taq polymerase gene fragment was replaced with reinforced, and a variety of chimeric DNA polymerases prepared. Based on these properties chimeric enzymes and their sequence, two residues, E742 and A743, in the Taq polymerase was found to be important for the ability of elongation. Taq polymerase with mutations in the 742 and 743 actually showed a higher affinity DNA and primer extension capabilities faster. These factors also affect the performance of PCR on DNA polymerase, and the PCR yield improvement was observed with Taq polymerase mutants. Fast-cycling PCR formulations, protocols, and instruments have been developed to address the need to improve throughput and shorter turn-around times for PCR-based tests.

Although the run time can be cut by up to 50% shorter cycle times have been correlated with a lower detection sensitivity and increased variability. To solve this problem, we apply the self compartmentalized Replication (CSR) to develop faster-cycling mutant Taq DNA polymerase. After five rounds of selection using a shorter PCR extension times, individual mutations identified in the fastest-cycling clones randomly combined using ligation-based multi-site mutagenesis.

The best performing combination mutants showed 35- 90-fold higher affinity (lower Kd) for excellent template and (2-fold) increase in the rate of extension moderate compared to wild-type Taq. Further characterization revealed that mutations selected CSR provides increased resistance to inhibitors, and in particular, enable direct amplification of up to 65% whole blood. We discuss the contribution of individual mutations for fast-cycling and blood-resistant phenotype.

Mutant Taq DNA polymerases with improved elongation ability as a useful reagent for genetic engineering.
Mutant Taq DNA polymerases with improved elongation ability as a useful reagent for genetic engineering.

The increase in the gap repair cloning in yeast: vector treatment gap with Taq DNA polymerase Avoid self-ligation of the vector.

Gap repair a fast and efficient method for the assembly of recombinant DNA molecules in Saccharomyces cerevisiae. This method produces a circular DNA molecule by homologous recombination between two or more linear DNA fragments, one of which is usually the vector carrying the replicative sequence and a selective marker.

This technique avoids laborious and expensive in vitro purification and DNA ligation. DNA repair machinery can also close and ligate linear vector by a mechanism other than homologous recombination, resulting in an empty vector. The frequency of adverse events can be reduced by removing the 5′-phosphate group using a phosphatase, which is the standard method used for in vitro ligation.

Taq DNA polymerase
BT10102 1000U
EUR 130.8
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA polymerase
BT10103 2000U
EUR 200.4
Description: High quality Taq polymerase for different PCR variations and downstream applications.

Taq DNA Polymerase
9001-2500 each
EUR 614.4

Taq DNA Polymerase
9001-500 each
EUR 229.2

Taq DNA Polymerase
L7051001 1000 unit
EUR 159

Taq DNA Polymerase
L7051200 200 unit
EUR 41

Taq DNA Polymerase (3000U)
9K-001-0002 3000U
EUR 582

Taq DNA Polymerase (6000U)
9K-001-0033 6000U
EUR 1234.5

Taq DNA Polymerase (1000U)
9K-001-0035 1000U
EUR 418.62

Taq DNA Polymerase (500U)
9K-001-0001 500U
EUR 209.29

Taq HS DNA Polymerase
P132-d1 500 U
EUR 38.28

Taq HS DNA Polymerase
P132-d2 1,000 U
EUR 71.34

Taq HS DNA Polymerase
P132-d3 5,000 U
EUR 324.8

Taq DNA Polymerase (PCR)
G008 5000 U (1.0 ml)
EUR 246

Taq DNA Polymerase (PCR)
G009 1000 U (200 ul)
EUR 104.4

Taq DNA Polymerase (PCR)
G126 10000 U (2 x 1.0 ml)
EUR 370.8

Taq DNA polymerase (+dNTPs)
02-001 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 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 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 5×200U
EUR 261.6
Description: The Taq DNA polymerase (-dNTPs) is available in Europe and for worldwide shipping via Gentaur.

Maximo Taq DNA Polymerase
S101 500 units
EUR 55.2

Maximo Taq DNA Polymerase
S102 5x500 units
EUR 128.4

Maximo Taq DNA Polymerase
S103 20x500 units
EUR 393.6

Taq DNA Polymerase, 5units/㎕
T2200-050 500units
EUR 157.2

Taq DNA Polymerase, 5units/㎕
T2200-100 2x500 units
EUR 230.4

Taq DNA Polymerase, 5units/㎕
T2200-200 4x500 units
EUR 375.6

Hotstart Taq DNA Polymerase
L7052001 1000 units
EUR 567

Hotstart Taq DNA Polymerase
L7052250 250 units
EUR 159

Champagne Taq DNA Polymerase
P122-d1 500 U (2.5 U/μl)
EUR 212.4

Champagne Taq DNA Polymerase
P122-d2 500 U (5 U/μl)
EUR 212.4

Champagne Taq DNA Polymerase
P122-d3 500 U (10 U/μl)
EUR 212.4

ACTaq? Taq DNA Polymerase, 1000U
E2100-1000U each
EUR 260.4

Taq DNA Polymerase - 5,000 units
3245 1/EA
EUR 620.4

Recombinant Taq Dna Polymerase
27079804 EACH
EUR 117.6

Recombinant Taq Dna Polymerase
27079806 EACH
EUR 639.6

Taq DNA Polymerase (Recombinant)
abx073521-10000U 10000 U
EUR 710.4

Taq DNA Polymerase (Recombinant)
abx073521-1000U 1000 U
EUR 276

Taq DNA Polymerase (Recombinant)
abx073521-3000U 3000 U
EUR 393.6

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

ACTaq? Taq DNA Polymerase, 250U
E2100-250U each
EUR 122.4

Taq Plus DNA Polymerase
P201-01 250 U
EUR 147.6

Taq Plus DNA Polymerase
P201-02 1000 U
EUR 213.6

Taq Plus DNA Polymerase
P201-03 3000 U
EUR 382.8

Taq DNA Polymerase, 5u/ul
B0089(D0089) 5X200U
EUR 112.2

Genorise® Taq DNA Polymerase
GR108007 200 tests
EUR 230

YEA taq DNA Polymerase wiodNTP
FYT011-500U 500U Ask for price

Taq Plus DNA Polymerase (PCR)
G012 250 U (50 ul)
EUR 87.6

Taq Plus DNA Polymerase (PCR)
G040 1000 U (200 ul)
EUR 127.2

Recombinant (E.Coli) Taq DNA Polymerase
RP-394 1000 IU
EUR 196.8

Taq DNA Polymerase Polyclonal Antibody
7018-100 each
EUR 424.8

Taq DNA Polymerase Polyclonal Antibody
7018-30T each
EUR 175.2

Taq DNA Polymerase, 5 units/µl
ABP-PP-ERTQ001 1000U Ask for price

Taq DNA Polymerase, 5 units/µl
ABP-PP-ERTQ100 100U Ask for price

Taq DNA Polymerase, 5 units/µl
ABP-PP-ERTQ200 200U Ask for price

Taq DNA Polymerase, 5 units/µl
ABP-PP-ERTQ500 500U Ask for price

YEA taq DNA Polymerase wi dNTP
FYT001-500U 500U Ask for price

ACTaq? Taq Blue DNA Polymerase, 1000U
E2100B-1000U each
EUR 272.4

Taq DNA Polymerase (Mg2+ plus Buffer)
P101-01 1000 U
EUR 145.2

Taq DNA Polymerase (Mg2+ plus Buffer)
P101-02 5000 U
EUR 231.6

Taq DNA Polymerase (Mg2+ plus Buffer)
P101-03 10000 U
EUR 314.4

Taq DNA Polymerase (Mg2+ free Buffer)
P102-01 1000 U
EUR 145.2

Taq DNA Polymerase (Mg2+ free Buffer)
P102-02 5000 U
EUR 231.6

Taq DNA Polymerase (Mg2+ free Buffer)
P102-03 10000 U
EUR 314.4

ACTaq? Taq Blue DNA Polymerase, 250U
E2100B-250U each
EUR 126

Taq DNA polymerase (TQ21) , DNA Aptamer, unlabeled
AD-146-U Custom Ask for price

Cheetah Hot Start Taq DNA Polymerase
29050 500EU
EUR 355.2
Description: Minimum order quantity: 1 unit of 500EU

Taq Plus DNA Polymerase, 5u/ul
D0090 200U
EUR 94.8

Taq Plus DNA Polymerase (Recombinant)
abx073522-2500U 2.500 U
EUR 710.4

Taq Plus DNA Polymerase (Recombinant)
abx073522-250U 250 U
EUR 276

Taq Plus DNA Polymerase (Recombinant)
abx073522-750U 750 U
EUR 393.6

Recombinant Taq Plus DNA Polymerase
7-03631 250U Ask for price

Recombinant Taq Plus DNA Polymerase
7-03632 750U Ask for price

Recombinant Taq Plus DNA Polymerase
7-03633 2,500U Ask for price

Taq HS DNA Polymerase (Glycerol-free)
QL101-01 1,000 U
EUR 71.2

Taq HS DNA Polymerase (Glycerol-free)
QL101-02 5,000 U
EUR 324.8

Taq HS DNA Polymerase (Glycerol-free)
QL101-03 50,000 U
EUR 2900

Taq DNA Polymerase, 5units/㎕ , Bulk Packing
T2200-B01 10,000 units
EUR 1388.4

Taq DNA Polymerase, 5units/㎕ , Bulk Packing
T2200-B05 5x10,000 units
EUR 6540

Taq DNA polymerase (TQ21) , DNA Aptamer, Biotinylated
AD-146-B Custom Ask for price

amfiSure Hot-Start Taq DNA Polymerase
P0322-010 100 units
EUR 176.4

amfiSure Hot-Start Taq DNA Polymerase
P0322-025 250 units
EUR 273.6

3G HotStart Taq DNA Polymerase (5u/ul)
3GHST81 5x500U
EUR 896.24

Taq DNA Polymerase with dNTP Mix (6000U)
9K-001-0018 6000U
EUR 1536.74

Taq DNA Polymerase with dNTP Mix (500U)
9K-001-0031 500U
EUR 351.28

Taq DNA Polymerase with dNTP Mix (3000U)
9K-001-0032 3000U
EUR 1088.87

Taq DNA Polymerase with dNTP Mix (1000U)
9K-001-0034 1000U
EUR 518.84

Taq DNA Polymerase (High Purity), 5u/ul
HTD0078 5X200U
EUR 129.6

HiQTM Taq Red DNA Polymerase, 1unit/㎕
T2201-050 500units
EUR 157.2

HiQTM Taq Red DNA Polymerase, 1unit/㎕
T2201-100 2x500 units
EUR 230.4

HiQTM Taq Red DNA Polymerase, 1unit/㎕
T2201-200 4x500 units
EUR 375.6

Genorise® CLT Taq DNA Polymerase (1000 U)
GR108003 1000 U
EUR 199

Maximo DFS-Plus Taq DNA Polymerase DNA-free
N140 500 units
EUR 60

Maximo DFS-Plus Taq DNA Polymerase DNA-free
N142 5x500 units
EUR 144

Maximo DFS-Plus Taq DNA Polymerase DNA-free
N144 20x500 units
EUR 440.4

Taq DNA polymerase (TQ21) , DNA Aptamer, FITC labeled
AD-146-F Custom Ask for price

Taq DNA Polymerase 2x Premix - 500 Reactions
3249 1/EA
EUR 228

HotStart Taq DNA Polymerase (Mg2+ Plus Buffer)
RK20601 25 U Ask for price

Recombinant (E.Coli) Taq Plus DNA Polymerase
RP-395 250 U
EUR 196.8

Taq DNA Polymerase, Ultra Pure, 5 units/ul
T2016-001 1000 units
EUR 198

Taq DNA Polymerase, Ultra Pure, 5 units/ul
T2016-005 5000 units
EUR 360

Taq DNA Polymerase, Ultra Pure, 5 units/ul
T2016-010 10,000 units
EUR 699.6

Taq DNA Polymerase, Ultra Pure, 5 units/ul
T2016-100 100,000 units
EUR 3462

Fast-Taq DNA Polymerase (500U)
9K-001-0036 500U
EUR 420.65

amfiXpand Taq DNA Polymerase,Including dNTP&MgCl2
P0330-010 100 units
EUR 180

amfiXpand Taq DNA Polymerase,Including dNTP&MgCl3
P0330-020 2x100 units
EUR 286.8

amfiXpand Taq DNA Polymerase,Including dNTP&MgCl2
P0330-050 5X100 units
EUR 465.6

amfiXpand Taq DNA Polymerase,Including dNTP&MgCl2
P0330-250 25X100 units
EUR 2004

amfiXpand Taq DNA Polymerase,Including dNTP&MgCl2
P0330-500 50X100 units
EUR 3591.6

Taq DNA Polymerase (Mg2+ plus Buffer, with dNTP)
P101-d1 1000 U
EUR 163.2

Taq DNA Polymerase (Mg2+ plus Buffer, with dNTP)
P101-d2 5000 U
EUR 320.4

Taq DNA Polymerase (Mg2+ plus Buffer, with dNTP)
P101-d3 10000 U
EUR 490.8

Taq DNA Polymerase (Mg2+ free Buffer, with dNTP)
P102-d1 1000 U
EUR 163.2

Taq DNA Polymerase (Mg2+ free Buffer, with dNTP)
P102-d2 5000 U
EUR 320.4

Taq DNA Polymerase (Mg2+ free Buffer, with dNTP)
P102-d3 10000 U
EUR 490.8

Reversible 1-min Hot Start Taq DNA Polymerase
P950 - Ask for price

Reversible 1-min Hot Start Taq DNA Polymerase
P950L - Ask for price

Taq DNA Polymerase - 5,000 units with separate dNTP Mix
3245d 1/EA
EUR 802.8

Anti-Taq DNA Polymerase Antibody
48369-100ul 100ul
EUR 399.6

Anti-Taq DNA Polymerase Antibody
48369-50ul 50ul
EUR 286.8

DFS-"HOT" Taq DNA Polymerase (DNA-free sensitive, E-coli. DNA Free)
N150 500 units
EUR 122.4

DFS-"HOT" Taq DNA Polymerase (DNA-free sensitive, E-coli. DNA Free)
N152 5x500 units
EUR 472.8

DFS-"HOT" Taq DNA Polymerase (DNA-free sensitive, E-coli. DNA Free)
N154 20x500 units
EUR 1830

Taq Pro Multiplex DNA Polymerase (High sensitivity)
PM201-01 200 rxns
EUR 72.5

Taq Pro Multiplex DNA Polymerase (High sensitivity)
PM201-02 1,000 rxns
EUR 290

Taq Pro Multiplex DNA Polymerase (High specificity)
PM202-01 200 rxns
EUR 72.5

Taq Pro Multiplex DNA Polymerase (High specificity)
PM202-02 1,000 rxns
EUR 290

HiQTM Taq Red DNA Polymerase, 1unit/㎕ , Bulk Packing
T2201-B01 10,000 units
EUR 1388.4

HiQTM Taq Red DNA Polymerase, 1unit/㎕ , Bulk Packing
T2201-B05 5x10,000 units
EUR 6540

Hot Start Taq DNA Polymerase - 1000 units with seperate dNTP mix
3293d 1/EA
EUR 428.4

amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2
P0310-025 250 unit
EUR 200.4

amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2
P0310-050 2X250 unit
EUR 300

amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2
P0310-100 4X250 Unit
EUR 505.2

amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2
P0310-250 10X250 Unit
EUR 886.8

Maximo Taq DNA Polymerase (2X pre-mix, ready-to-use)
S113 2x100 rcs
EUR 79.2

Maximo Taq DNA Polymerase (2X pre-mix, ready-to-use)
S114 10x100 rcs
EUR 231.6

Maximo H-SPlus Taq DNA Polymerase (for Hot-Start PCR)
S400 200 units
EUR 86.4

Maximo H-SPlus Taq DNA Polymerase (for Hot-Start PCR)
S410 1000 units
EUR 231.6

amfiEco Taq DNA Polymerase, 1 unit/ul,Including dNTP&MgCl2
P0701-050 500 units
EUR 198

amfiEco Taq DNA Polymerase, 1 unit/ul,Including dNTP&MgCl2
P0701-102 25x500 units
EUR 1712.4

amfiEco Taq DNA Polymerase, 1 unit/ul,Including dNTP&MgCl2
P0701-500 10X500 units
EUR 886.8

Anti-Taq DNA Polymerase Conjugated Antibody
C48369 100ul
EUR 476.4

amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-500 500 units
EUR 262.8

amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-502 2x500 units
EUR 424.8

amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-510 10x500 units
EUR 1647.6

amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-525 25x500 units
EUR 3624

amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-550 50x500 units
EUR 6459.6

EZ Taq Polymerase (HotStart)
9K-005-0020 500U
EUR 245.83

Accuris Taq Polymerase, 1000 units
PR1000-1000 1 PC
EUR 167.1

Accuris Taq Polymerase, 6000 units
PR1000-6000 1 PC
EUR 853.3

Accuris Taq Polymerase, 500 units
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)
9K-001-0020 5000U
EUR 2878.8

High-Taq DNA Polymerase with 10mM dNTP Mix (4x250U)
9K-001-0006 4x250U
EUR 1055.98

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

Pfu DNA Polymerase
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

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

Ready? DNA Polymerase
M1146-1000 each
EUR 398.4

Ready? DNA Polymerase
M1146-10000 each
EUR 1279.2

Ready? DNA Polymerase
M1146-5000 each
EUR 960

Phi29 DNA Polymerase
M1239-1000 each
EUR 373.2

Kodaq DNA Polymerase
G498 250 U (50 ul)
EUR 104.4

However, phosphatase treatment is less effective for gap repair cloning rather than in vitro ligation, probably due to the ability of S. cerevisiae DNA repair machinery to efficiently repair the lost phosphate groups to allow religation. We have developed a more efficient method to prevent vector religation, the vector fragment by treatment with Taq DNA polymerase and dATP. It prevents the procedure recircularization vector almost completely, to facilitate the screening of recombinant clones correctly.

Jordan

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