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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.
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
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
Taq DNA Polymerase
L7051001
Biochain
1000 unit
EUR 159
Taq DNA Polymerase
L7051200
Biochain
200 unit
EUR 41
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
Taq DNA Polymerase (500U)
9K-001-0001
Bio Basic
500U
EUR 209.29
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
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
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.
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
Hotstart Taq DNA Polymerase
L7052001
Biochain
1000 units
EUR 567
Hotstart Taq DNA Polymerase
L7052250
Biochain
250 units
EUR 159
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
ACTaq? Taq DNA Polymerase, 1000U
E2100-1000U
ACTGene
each
EUR 260.4
Taq DNA Polymerase (Recombinant)
abx073521-10000U
Abbexa
10000 U
EUR 710.4
Taq DNA Polymerase (Recombinant)
abx073521-1000U
Abbexa
1000 U
EUR 276
Taq DNA Polymerase (Recombinant)
abx073521-3000U
Abbexa
3000 U
EUR 393.6
Recombinant Taq DNA Polymerase
7-03634
CHI Scientific
1,000U
Ask for price
Recombinant Taq DNA Polymerase
7-03635
CHI Scientific
3,000U
Ask for price
Recombinant Taq DNA Polymerase
7-03636
CHI Scientific
10,000U
Ask for price
ACTaq? Taq DNA Polymerase, 250U
E2100-250U
ACTGene
each
EUR 122.4
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
Taq DNA Polymerase, 5u/ul
B0089(D0089)
Bio Basic
5X200U
EUR 112.2
Taq Plus DNA Polymerase (PCR)
G012
ABM
250 U (50 ul)
EUR 87.6
Taq Plus DNA Polymerase (PCR)
G040
ABM
1000 U (200 ul)
EUR 127.2
Taq DNA Polymerase Polyclonal Antibody
7018-100
Biovision
each
EUR 424.8
Taq DNA Polymerase Polyclonal Antibody
7018-30T
Biovision
each
EUR 175.2
Taq DNA Polymerase, 5 units/µl
ABP-PP-ERTQ001
Allele Biotech
1000U
Ask for price
Taq DNA Polymerase, 5 units/µl
ABP-PP-ERTQ100
Allele Biotech
100U
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Taq DNA Polymerase, 5 units/µl
ABP-PP-ERTQ200
Allele Biotech
200U
Ask for price
Taq DNA Polymerase, 5 units/µl
ABP-PP-ERTQ500
Allele Biotech
500U
Ask for price
ACTaq? Taq Blue DNA Polymerase, 1000U
E2100B-1000U
ACTGene
each
EUR 272.4
Taq DNA Polymerase (Mg2+ plus Buffer)
P101-01
Vazyme
1000 U
EUR 145.2
Taq DNA Polymerase (Mg2+ plus Buffer)
P101-02
Vazyme
5000 U
EUR 231.6
Taq DNA Polymerase (Mg2+ plus Buffer)
P101-03
Vazyme
10000 U
EUR 314.4
Taq DNA Polymerase (Mg2+ free Buffer)
P102-01
Vazyme
1000 U
EUR 145.2
Taq DNA Polymerase (Mg2+ free Buffer)
P102-02
Vazyme
5000 U
EUR 231.6
Taq DNA Polymerase (Mg2+ free Buffer)
P102-03
Vazyme
10000 U
EUR 314.4
ACTaq? Taq Blue DNA Polymerase, 250U
E2100B-250U
ACTGene
each
EUR 126
Taq DNA polymerase (TQ21) , DNA Aptamer, unlabeled
AD-146-U
Alpha Diagnostics
Custom
Ask for price
Cheetah Hot Start Taq DNA Polymerase
29050
Biotium
500EU
EUR 355.2
Description: Minimum order quantity: 1 unit of 500EU
Taq Plus DNA Polymerase, 5u/ul
D0090
Bio Basic
200U
EUR 94.8
Taq Plus DNA Polymerase (Recombinant)
abx073522-2500U
Abbexa
2.500 U
EUR 710.4
Taq Plus DNA Polymerase (Recombinant)
abx073522-250U
Abbexa
250 U
EUR 276
Taq Plus DNA Polymerase (Recombinant)
abx073522-750U
Abbexa
750 U
EUR 393.6
Recombinant Taq Plus DNA Polymerase
7-03631
CHI Scientific
250U
Ask for price
Recombinant Taq Plus DNA Polymerase
7-03632
CHI Scientific
750U
Ask for price
Recombinant Taq Plus DNA Polymerase
7-03633
CHI Scientific
2,500U
Ask for price
Taq HS DNA Polymerase (Glycerol-free)
QL101-01
Vazyme
1,000 U
EUR 71.2
Taq HS DNA Polymerase (Glycerol-free)
QL101-02
Vazyme
5,000 U
EUR 324.8
Taq HS DNA Polymerase (Glycerol-free)
QL101-03
Vazyme
50,000 U
EUR 2900
Taq DNA Polymerase, 5units/㎕ , Bulk Packing
T2200-B01
GenDepot
10,000 units
EUR 1388.4
Taq DNA Polymerase, 5units/㎕ , Bulk Packing
T2200-B05
GenDepot
5x10,000 units
EUR 6540
Taq DNA polymerase (TQ21) , DNA Aptamer, Biotinylated
AD-146-B
Alpha Diagnostics
Custom
Ask for price
amfiSure Hot-Start Taq DNA Polymerase
P0322-010
GenDepot
100 units
EUR 176.4
amfiSure Hot-Start Taq DNA Polymerase
P0322-025
GenDepot
250 units
EUR 273.6
3G HotStart Taq DNA Polymerase (5u/ul)
3GHST81
Bio Basic
5x500U
EUR 896.24
Taq DNA Polymerase with dNTP Mix (6000U)
9K-001-0018
Bio Basic
6000U
EUR 1536.74
Taq DNA Polymerase with dNTP Mix (500U)
9K-001-0031
Bio Basic
500U
EUR 351.28
Taq DNA Polymerase with dNTP Mix (3000U)
9K-001-0032
Bio Basic
3000U
EUR 1088.87
Taq DNA Polymerase with dNTP Mix (1000U)
9K-001-0034
Bio Basic
1000U
EUR 518.84
Taq DNA Polymerase (High Purity), 5u/ul
HTD0078
Bio Basic
5X200U
EUR 129.6
HiQTM Taq Red DNA Polymerase, 1unit/㎕
T2201-050
GenDepot
500units
EUR 157.2
HiQTM Taq Red DNA Polymerase, 1unit/㎕
T2201-100
GenDepot
2x500 units
EUR 230.4
HiQTM Taq Red DNA Polymerase, 1unit/㎕
T2201-200
GenDepot
4x500 units
EUR 375.6
Maximo DFS-Plus Taq DNA Polymerase DNA-free
N140
GeneOn
500 units
EUR 60
Maximo DFS-Plus Taq DNA Polymerase DNA-free
N142
GeneOn
5x500 units
EUR 144
Maximo DFS-Plus Taq DNA Polymerase DNA-free
N144
GeneOn
20x500 units
EUR 440.4
Taq DNA polymerase (TQ21) , DNA Aptamer, FITC labeled
AD-146-F
Alpha Diagnostics
Custom
Ask for price
Taq DNA Polymerase 2x Premix - 500 Reactions
3249
Intact Genomics
1/EA
EUR 228
HotStart Taq DNA Polymerase (Mg2+ Plus Buffer)
RK20601
Abclonal
25 U
Ask for price
Recombinant (E.Coli) Taq Plus DNA Polymerase
RP-395
Alpha Diagnostics
250 U
EUR 196.8
Taq DNA Polymerase, Ultra Pure, 5 units/ul
T2016-001
GenDepot
1000 units
EUR 198
Taq DNA Polymerase, Ultra Pure, 5 units/ul
T2016-005
GenDepot
5000 units
EUR 360
Taq DNA Polymerase, Ultra Pure, 5 units/ul
T2016-010
GenDepot
10,000 units
EUR 699.6
Taq DNA Polymerase, Ultra Pure, 5 units/ul
T2016-100
GenDepot
100,000 units
EUR 3462
Fast-Taq DNA Polymerase (500U)
9K-001-0036
Bio Basic
500U
EUR 420.65
amfiXpand Taq DNA Polymerase,Including dNTP&MgCl2
P0330-010
GenDepot
100 units
EUR 180
amfiXpand Taq DNA Polymerase,Including dNTP&MgCl3
P0330-020
GenDepot
2x100 units
EUR 286.8
amfiXpand Taq DNA Polymerase,Including dNTP&MgCl2
P0330-050
GenDepot
5X100 units
EUR 465.6
amfiXpand Taq DNA Polymerase,Including dNTP&MgCl2
P0330-250
GenDepot
25X100 units
EUR 2004
amfiXpand Taq DNA Polymerase,Including dNTP&MgCl2
P0330-500
GenDepot
50X100 units
EUR 3591.6
Taq DNA Polymerase (Mg2+ plus Buffer, with dNTP)
P101-d1
Vazyme
1000 U
EUR 163.2
Taq DNA Polymerase (Mg2+ plus Buffer, with dNTP)
P101-d2
Vazyme
5000 U
EUR 320.4
Taq DNA Polymerase (Mg2+ plus Buffer, with dNTP)
P101-d3
Vazyme
10000 U
EUR 490.8
Taq DNA Polymerase (Mg2+ free Buffer, with dNTP)
P102-d1
Vazyme
1000 U
EUR 163.2
Taq DNA Polymerase (Mg2+ free Buffer, with dNTP)
P102-d2
Vazyme
5000 U
EUR 320.4
Taq DNA Polymerase (Mg2+ free Buffer, with dNTP)
P102-d3
Vazyme
10000 U
EUR 490.8
Reversible 1-min Hot Start Taq DNA Polymerase
P950
101Bio
-
Ask for price
Reversible 1-min Hot Start Taq DNA Polymerase
P950L
101Bio
-
Ask for price
Taq DNA Polymerase - 5,000 units with separate dNTP Mix
3245d
Intact Genomics
1/EA
EUR 802.8
Anti-Taq DNA Polymerase Antibody
48369-100ul
SAB
100ul
EUR 399.6
Anti-Taq DNA Polymerase Antibody
48369-50ul
SAB
50ul
EUR 286.8
DFS-"HOT" Taq DNA Polymerase (DNA-free sensitive, E-coli. DNA Free)
N150
GeneOn
500 units
EUR 122.4
DFS-"HOT" Taq DNA Polymerase (DNA-free sensitive, E-coli. DNA Free)
N152
GeneOn
5x500 units
EUR 472.8
DFS-"HOT" Taq DNA Polymerase (DNA-free sensitive, E-coli. DNA Free)
N154
GeneOn
20x500 units
EUR 1830
Taq Pro Multiplex DNA Polymerase (High sensitivity)
PM201-01
Vazyme
200 rxns
EUR 72.5
Taq Pro Multiplex DNA Polymerase (High sensitivity)
PM201-02
Vazyme
1,000 rxns
EUR 290
Taq Pro Multiplex DNA Polymerase (High specificity)
PM202-01
Vazyme
200 rxns
EUR 72.5
Taq Pro Multiplex DNA Polymerase (High specificity)
PM202-02
Vazyme
1,000 rxns
EUR 290
HiQTM Taq Red DNA Polymerase, 1unit/㎕ , Bulk Packing
T2201-B01
GenDepot
10,000 units
EUR 1388.4
HiQTM Taq Red DNA Polymerase, 1unit/㎕ , Bulk Packing
T2201-B05
GenDepot
5x10,000 units
EUR 6540
Hot Start Taq DNA Polymerase - 1000 units with seperate dNTP mix
3293d
Intact Genomics
1/EA
EUR 428.4
amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2
P0310-025
GenDepot
250 unit
EUR 200.4
amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2
P0310-050
GenDepot
2X250 unit
EUR 300
amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2
P0310-100
GenDepot
4X250 Unit
EUR 505.2
amfiSure Taq DNA Polymerase, 1unit/ul, Including dNTP&MgCl2
P0310-250
GenDepot
10X250 Unit
EUR 886.8
Maximo Taq DNA Polymerase (2X pre-mix, ready-to-use)
S113
GeneOn
2x100 rcs
EUR 79.2
Maximo Taq DNA Polymerase (2X pre-mix, ready-to-use)
S114
GeneOn
10x100 rcs
EUR 231.6
Maximo H-SPlus Taq DNA Polymerase (for Hot-Start PCR)
S400
GeneOn
200 units
EUR 86.4
Maximo H-SPlus Taq DNA Polymerase (for Hot-Start PCR)
S410
GeneOn
1000 units
EUR 231.6
amfiEco Taq DNA Polymerase, 1 unit/ul,Including dNTP&MgCl2
P0701-050
GenDepot
500 units
EUR 198
amfiEco Taq DNA Polymerase, 1 unit/ul,Including dNTP&MgCl2
P0701-102
GenDepot
25x500 units
EUR 1712.4
amfiEco Taq DNA Polymerase, 1 unit/ul,Including dNTP&MgCl2
P0701-500
GenDepot
10X500 units
EUR 886.8
Anti-Taq DNA Polymerase Conjugated Antibody
C48369
SAB
100ul
EUR 476.4
amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-500
GenDepot
500 units
EUR 262.8
amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-502
GenDepot
2x500 units
EUR 424.8
amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-510
GenDepot
10x500 units
EUR 1647.6
amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-525
GenDepot
25x500 units
EUR 3624
amfiSure ONE Taq DNA Polymerase, 1unit/ul, MgCl2 and dNTP Included
P7002-550
GenDepot
50x500 units
EUR 6459.6
EZ Taq Polymerase (HotStart)
9K-005-0020
Bio Basic
500U
EUR 245.83
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 with 10mM dNTP Mix (500U)
9K-001-0003
Bio Basic
500U
EUR 575.22
Fast-Taq DNA Polymerase with 10mM dNTP Mix (2500U)
9K-001-0004
Bio Basic
2500U
EUR 2465.38
High-Taq DNA Polymerase with 10mM dNTP Mix (250U)
9K-001-0005
Bio Basic
250U
EUR 384.17
High-Taq DNA Polymerase with 10mM dNTP Mix (5000U)
9K-001-0020
Bio Basic
5000U
EUR 2878.8
High-Taq DNA Polymerase with 10mM dNTP Mix (4x250U)
9K-001-0006
Bio Basic
4x250U
EUR 1055.98
T4 DNA Polymerase
N101-01
Vazyme
2000 U
EUR 278.4
T4 DNA Polymerase
M1211-100
Biovision
each
EUR 385.2
DNA polymerase ? (Rat)
10-101
Sceti
20ug
EUR 465.6
Description: The DNA polymerase ? (Rat) is available in Europe and for worldwide shipping via Gentaur.
DNA polymerase ? (Rat)
10-102
Sceti
100ug
EUR 1107.6
Description: The DNA polymerase ? (Rat) is available in Europe and for worldwide shipping via Gentaur.
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.
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
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
Ask for price
MyFi DNA Polymerase
BIO-21117/S
Bioline
Sample
Ask for price
MyFi DNA Polymerase
BIO-21118
Bioline
500 Units
Ask for price
MyFi DNA Polymerase
BIO-21119
Bioline
2500 Units
Ask for price
MyTaq DNA Polymerase
BIO-21105
Bioline
500 units
Ask for price
MyTaq DNA Polymerase
BIO-21105/S
Bioline
Sample
Ask for price
MyTaq DNA Polymerase
BIO-21106
Bioline
2500 units
Ask for price
MyTaq DNA Polymerase
BIO-21107
Bioline
5000 units
Ask for price
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
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.