Structured Review

TaKaRa taq polymerase
Optimization of the <t>UP-M-PCR.</t> Lane A, B, C, D, E, amplicon fragments by UP (500 nmol L −1 ) and compound specific primer hpt-839, nptII-508, pat-262, bar-226 and sps-110 at a series concentrations of 500 nmol L −1 , 50 nmol L −1 , 25 nmol L −1 , 5 nmol L −1 , 0.5 nmol L −1 ; lane F1, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at 25 nmol L −1 ; lane F2, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at the optimized concentration; lane G1,G2,G3, amplicon fragments by all primers at the optimized concentration with TaKaRa <t>Taq</t> ™, Phire™ Hot Start DNA polymerase, iProof™ High-Fidelity DNA polymerase; lane H1, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the common amplification conditions; lane H2, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the optimized amplification conditions; lane M, 100 bp DNA Marker.
Taq Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 1138 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/taq polymerase/product/TaKaRa
Average 92 stars, based on 1138 article reviews
Price from $9.99 to $1999.99
taq polymerase - by Bioz Stars, 2020-05
92/100 stars

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1) Product Images from "A Novel Universal Primer-Multiplex-PCR Method with Sequencing Gel Electrophoresis Analysis"

Article Title: A Novel Universal Primer-Multiplex-PCR Method with Sequencing Gel Electrophoresis Analysis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0022900

Optimization of the UP-M-PCR. Lane A, B, C, D, E, amplicon fragments by UP (500 nmol L −1 ) and compound specific primer hpt-839, nptII-508, pat-262, bar-226 and sps-110 at a series concentrations of 500 nmol L −1 , 50 nmol L −1 , 25 nmol L −1 , 5 nmol L −1 , 0.5 nmol L −1 ; lane F1, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at 25 nmol L −1 ; lane F2, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at the optimized concentration; lane G1,G2,G3, amplicon fragments by all primers at the optimized concentration with TaKaRa Taq ™, Phire™ Hot Start DNA polymerase, iProof™ High-Fidelity DNA polymerase; lane H1, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the common amplification conditions; lane H2, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the optimized amplification conditions; lane M, 100 bp DNA Marker.
Figure Legend Snippet: Optimization of the UP-M-PCR. Lane A, B, C, D, E, amplicon fragments by UP (500 nmol L −1 ) and compound specific primer hpt-839, nptII-508, pat-262, bar-226 and sps-110 at a series concentrations of 500 nmol L −1 , 50 nmol L −1 , 25 nmol L −1 , 5 nmol L −1 , 0.5 nmol L −1 ; lane F1, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at 25 nmol L −1 ; lane F2, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at the optimized concentration; lane G1,G2,G3, amplicon fragments by all primers at the optimized concentration with TaKaRa Taq ™, Phire™ Hot Start DNA polymerase, iProof™ High-Fidelity DNA polymerase; lane H1, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the common amplification conditions; lane H2, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the optimized amplification conditions; lane M, 100 bp DNA Marker.

Techniques Used: Polymerase Chain Reaction, Amplification, Concentration Assay, Marker

2) Product Images from "A Novel Universal Primer-Multiplex-PCR Method with Sequencing Gel Electrophoresis Analysis"

Article Title: A Novel Universal Primer-Multiplex-PCR Method with Sequencing Gel Electrophoresis Analysis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0022900

Optimization of the UP-M-PCR. Lane A, B, C, D, E, amplicon fragments by UP (500 nmol L −1 ) and compound specific primer hpt-839, nptII-508, pat-262, bar-226 and sps-110 at a series concentrations of 500 nmol L −1 , 50 nmol L −1 , 25 nmol L −1 , 5 nmol L −1 , 0.5 nmol L −1 ; lane F1, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at 25 nmol L −1 ; lane F2, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at the optimized concentration; lane G1,G2,G3, amplicon fragments by all primers at the optimized concentration with TaKaRa Taq ™, Phire™ Hot Start DNA polymerase, iProof™ High-Fidelity DNA polymerase; lane H1, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the common amplification conditions; lane H2, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the optimized amplification conditions; lane M, 100 bp DNA Marker.
Figure Legend Snippet: Optimization of the UP-M-PCR. Lane A, B, C, D, E, amplicon fragments by UP (500 nmol L −1 ) and compound specific primer hpt-839, nptII-508, pat-262, bar-226 and sps-110 at a series concentrations of 500 nmol L −1 , 50 nmol L −1 , 25 nmol L −1 , 5 nmol L −1 , 0.5 nmol L −1 ; lane F1, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at 25 nmol L −1 ; lane F2, amplicon fragments by UP at 500 nmol L −1 and all compound specific primers at the optimized concentration; lane G1,G2,G3, amplicon fragments by all primers at the optimized concentration with TaKaRa Taq ™, Phire™ Hot Start DNA polymerase, iProof™ High-Fidelity DNA polymerase; lane H1, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the common amplification conditions; lane H2, amplicon fragments by all primers at the optimized concentration with Phire™ Hot Start DNA polymerase under the optimized amplification conditions; lane M, 100 bp DNA Marker.

Techniques Used: Polymerase Chain Reaction, Amplification, Concentration Assay, Marker

Related Articles

Polymerase Chain Reaction:

Article Title: A Novel Universal Primer-Multiplex-PCR Method with Sequencing Gel Electrophoresis Analysis
Article Snippet: .. To test the efficiency of Taq Polymerase to be employed in PCR assays, comparative tests were made with several Taq polymerases, such as Phire™ Hot Start DNA polymerase, iProof™ High-Fidelity DNA polymerase, and TaKaRa Taq ™. .. The Phire™ HotStart DNA polymerase, coupled with a preoptimized primer mix for different multiplex reactions, gave the best results both in terms of reproducibility and robustness.

Article Title: Long-term ethanol exposure inhibits glucose transporter 4 expression via an AMPK-dependent pathway in adipocytes
Article Snippet: .. PCR amplification was carried out in a total reaction volume of 25 μL, including 2.5 μL PCR buffer (10×), 0.2 μL Taq polymerase, 2 μL dNTP (TaKaRa, 2.5 mmol/L), 2 μL MgCl2 (TaKaRa, 25 mmol/L), 2 μL primers (5×10−6 mol/L) and 2.5−3 μL of the cDNA (2.5 μL for AMPK α1, α2, and GLUT4; 3 μL for MEF2A). .. The PCR products were subjected to 1.5% agarose gel electrophoresis containing ethidium bromide and visualized by excitation under UV light, quantified using Alphaimager 2200.

Article Title: Development and Validation of PCR-RFLP Assay for Identification of Gambierdiscus species in the Greater Caribbean region
Article Snippet: .. PCR amplification reactions (25 μl) contained ~5 ng template DNA, 1× PCR Buffer (500 mM KCL and 100 mM Tris-HCl, pH 8.3), 0.25 mM of each dNTP, 0.5 μM of D1R primer, 0.5 μM of D2C primer, and 0.625 U of Taq DNA Polymerase (Takara Taq Bio Inc). .. Hot start PCR amplification was performed using a Eppendorf Mastercycler thermocycler following these conditions: 5 minutes denaturing at 94°C (after 1–2 minutes at 94°C the cycle was paused to add the Taq), followed by 35 cycles of 30 seconds denaturing at 94°C, 1 minute annealing at 50°C, 2 minutes elongation at 72°C, and a final elongation for 10 minutes at 72°C.

Nested PCR:

Article Title: Identification of Mycoplasma fermentans in Synovial Fluid Samples from Arthritis Patients with Inflammatory Disease
Article Snippet: .. When a nested PCR was to be performed, the first round used Taq polymerase from Pharmacia or, in some cases, TaKaRa Ex Taq , which contains a proofreading enzyme which increases the sensitivity of the reaction two- to threefold. ..

Amplification:

Article Title: Long-term ethanol exposure inhibits glucose transporter 4 expression via an AMPK-dependent pathway in adipocytes
Article Snippet: .. PCR amplification was carried out in a total reaction volume of 25 μL, including 2.5 μL PCR buffer (10×), 0.2 μL Taq polymerase, 2 μL dNTP (TaKaRa, 2.5 mmol/L), 2 μL MgCl2 (TaKaRa, 25 mmol/L), 2 μL primers (5×10−6 mol/L) and 2.5−3 μL of the cDNA (2.5 μL for AMPK α1, α2, and GLUT4; 3 μL for MEF2A). .. The PCR products were subjected to 1.5% agarose gel electrophoresis containing ethidium bromide and visualized by excitation under UV light, quantified using Alphaimager 2200.

Article Title: Development and Validation of PCR-RFLP Assay for Identification of Gambierdiscus species in the Greater Caribbean region
Article Snippet: .. PCR amplification reactions (25 μl) contained ~5 ng template DNA, 1× PCR Buffer (500 mM KCL and 100 mM Tris-HCl, pH 8.3), 0.25 mM of each dNTP, 0.5 μM of D1R primer, 0.5 μM of D2C primer, and 0.625 U of Taq DNA Polymerase (Takara Taq Bio Inc). .. Hot start PCR amplification was performed using a Eppendorf Mastercycler thermocycler following these conditions: 5 minutes denaturing at 94°C (after 1–2 minutes at 94°C the cycle was paused to add the Taq), followed by 35 cycles of 30 seconds denaturing at 94°C, 1 minute annealing at 50°C, 2 minutes elongation at 72°C, and a final elongation for 10 minutes at 72°C.

BAC Assay:

Article Title: A 100%-complete sequence reveals unusually simple genomic features in the hot-spring red alga Cyanidioschyzon merolae
Article Snippet: .. PCRs of BAC clones containing unsequenced regions were carried out using Taq polymerase with GC buffer (Takara LA Taq; Takara Bio Inc., Osaka, Japan) and specific primers complementary to sequences flanking the gaps. .. DNA walking annealing control primer technology (DNA Walking SpeedUp™ Kit; Seegene, Seoul, Korea) was also used to directly amplify unknown sequences adjacent to known sequences within a contig.

Clone Assay:

Article Title: A 100%-complete sequence reveals unusually simple genomic features in the hot-spring red alga Cyanidioschyzon merolae
Article Snippet: .. PCRs of BAC clones containing unsequenced regions were carried out using Taq polymerase with GC buffer (Takara LA Taq; Takara Bio Inc., Osaka, Japan) and specific primers complementary to sequences flanking the gaps. .. DNA walking annealing control primer technology (DNA Walking SpeedUp™ Kit; Seegene, Seoul, Korea) was also used to directly amplify unknown sequences adjacent to known sequences within a contig.

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  • 99
    TaKaRa la dna polymerase
    ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying <t>DNA</t> polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and <t>Takara</t> LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 .
    La Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/la dna polymerase/product/TaKaRa
    Average 99 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    la dna polymerase - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    99
    TaKaRa taq dna polymerase
    (A) Genotype in each generation of mutant mice confirmed by PCR analysis on isolated genomic <t>DNA.</t> PCR was performed on the isolated genomic DNA using <t>taq</t> DNA polymerase and primer sets (#1 and #2 are for wild-type and GluR2 delta7 KI, #3 and #4 are for
    Taq Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1599 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taq dna polymerase/product/TaKaRa
    Average 99 stars, based on 1599 article reviews
    Price from $9.99 to $1999.99
    taq dna polymerase - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    Image Search Results


    ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 .

    Journal: Scientific Reports

    Article Title: Amplification and next generation sequencing of near full-length human enteroviruses for identification and characterisation from clinical samples

    doi: 10.1038/s41598-018-30322-y

    Figure Lengend Snippet: ( A ) Gel electrophoresis of near full-length genome PCR products produced from four long amplifying DNA polymerases; KlenTaq LA (discontinued), AccuTaq LA, PrimeSTAR GXL and Takara LA Taq (separate gel) per manufacturer’s instructions for samples with high GC/secondary structures. M, HyperLadder 1 kb; 1, CVB3 Nancy; 2, CVB5 Faulkner; 3, H 2 O control. ( B ) Gel electrophoresis of near full-length genome PCR products produced from Takara LA Taq DNA polymerase. M, HyperLadder 1 kb; 1–4, known EV positives from NSW Health Pathology East virology diagnostic lab; 5, CVB3 Nancy; 6, H 2 .

    Article Snippet: First round PCR was performed by adding cDNA (5 µL) to a reaction mix (50 µL) containing Takara LA Buffer (1X), dNTPs (100 µM each), forward primer vir24 (0.2 µM), reverse primer vir20 (0.2 µM), nuclease-free water (29.5 µL) and Takara LA DNA polymerase (2.5 U).

    Techniques: Nucleic Acid Electrophoresis, Polymerase Chain Reaction, Produced, Diagnostic Assay

    (A) Genotype in each generation of mutant mice confirmed by PCR analysis on isolated genomic DNA. PCR was performed on the isolated genomic DNA using taq DNA polymerase and primer sets (#1 and #2 are for wild-type and GluR2 delta7 KI, #3 and #4 are for

    Journal: Neuroscience letters

    Article Title: Involvement of GluR2 and GluR3 subunit C-termini in the trigeminal spinal subnucleus caudalis and C1-C2 neurons in trigeminal neuropathic pain

    doi: 10.1016/j.neulet.2010.12.060

    Figure Lengend Snippet: (A) Genotype in each generation of mutant mice confirmed by PCR analysis on isolated genomic DNA. PCR was performed on the isolated genomic DNA using taq DNA polymerase and primer sets (#1 and #2 are for wild-type and GluR2 delta7 KI, #3 and #4 are for

    Article Snippet: PCR was performed on the isolated genomic DNA using taq DNA polymerase (TaKaRa Ex Taq™, Takara, Otsu, Japan) and primer sets (primer #21–23 for GluR2 delta7: #21, 5′-ACA GAG GAA GGT AGT GGA AGG GAG-3′; #22, 5′-CTT GGT TTG GTT GTT GGT CAT AGC-3′; #23, 5′-CTA GTG AAC CTC TTC GAG GGA C-3′; primer #31–33 for GluR3 delta7: #31, 5′-CCA ATA CTC CAC AGG GGC AAT TTA TC-3′; #32, 5′-CCG TTG ACT GTT TTG AAT CTC ACA CC-3′; #33, 5′-CTA GTG AAC CTC TTC GAG GGA C-3′).

    Techniques: Mutagenesis, Mouse Assay, Polymerase Chain Reaction, Isolation

    Blocking efficiency of three kinds of blocked primers in PCR amplifications mediated by Taq DNA polymerase and high-fidelity DNA polymerase. (a) Primer and template sets. WT and Mu indicate wild-type and mutant types, respectively. The 3'-blocked forward primer completely matches the WT sequence but forms a mismatch with the Mu sequence (Mu*1) at the 3'-end. The reverse primer matches both the WT and Mu sequences. The 3'-terminal nucleotide of the blocked primer is blocked with-Pi or-Amino C6, or replaced with ddCTP. (b) Blocking efficiency of-Pi,-Amino C6 or-ddC in PCR mediated by Taq DNA polymerase. (c) Blocking efficiency of-Pi,-Amino C6 or-ddC in PCR mediated by high-fidelity DNA polymerase (KOD FX). (d) Discrimination efficiency of the typical proofreading-PCR medicated by various amounts (0.5, 1, 1.5 and 2 U) of KOD FX DNA polymerase using the ddC-blocked primer. All PCR amplifications were performed in a total volume of 20 μL containing 1 U of Taq or KOD FX DNA polymerase (Panel b and c) or various amounts of KOD FX DNA polymerase (Panel d). The cycling conditions consisted of pre-denaturation at 94°C for 2 min, followed by 30 cycles of denaturation at 94°C for 20 s, annealing at 56°C for 20 s and extension at 72°C for 25 s. NTC: no-template control.

    Journal: PLoS ONE

    Article Title: Modified Proofreading PCR for Detection of Point Mutations, Insertions and Deletions Using a ddNTP-Blocked Primer

    doi: 10.1371/journal.pone.0123468

    Figure Lengend Snippet: Blocking efficiency of three kinds of blocked primers in PCR amplifications mediated by Taq DNA polymerase and high-fidelity DNA polymerase. (a) Primer and template sets. WT and Mu indicate wild-type and mutant types, respectively. The 3'-blocked forward primer completely matches the WT sequence but forms a mismatch with the Mu sequence (Mu*1) at the 3'-end. The reverse primer matches both the WT and Mu sequences. The 3'-terminal nucleotide of the blocked primer is blocked with-Pi or-Amino C6, or replaced with ddCTP. (b) Blocking efficiency of-Pi,-Amino C6 or-ddC in PCR mediated by Taq DNA polymerase. (c) Blocking efficiency of-Pi,-Amino C6 or-ddC in PCR mediated by high-fidelity DNA polymerase (KOD FX). (d) Discrimination efficiency of the typical proofreading-PCR medicated by various amounts (0.5, 1, 1.5 and 2 U) of KOD FX DNA polymerase using the ddC-blocked primer. All PCR amplifications were performed in a total volume of 20 μL containing 1 U of Taq or KOD FX DNA polymerase (Panel b and c) or various amounts of KOD FX DNA polymerase (Panel d). The cycling conditions consisted of pre-denaturation at 94°C for 2 min, followed by 30 cycles of denaturation at 94°C for 20 s, annealing at 56°C for 20 s and extension at 72°C for 25 s. NTC: no-template control.

    Article Snippet: The results showed that the use of 0.1–0.15 U of high-fidelity DNA polymerase combined with 1 U of Taq DNA polymerase in per 20 μL reaction resulted in the best discrimination and amplification effects ( ).

    Techniques: Blocking Assay, Polymerase Chain Reaction, Mutagenesis, Sequencing

    Optimization of annealing temperature and amount of high-fidelity DNA polymerase in the modified PR-PCR. (a) Discrimination efficiency of the modified PR-PCR for known mutations using a mixture of 1 U of Taq DNA polymerase and various amount (0.05, 0.1 and 0.3 U) of KOD FX DNA polymerase in the reaction. The PCR amplifications were performed in a total volume of 20 μL under cycling conditions consisting of pre-denaturation at 94°C for 2 min, followed by 30 cycles of denaturation at 94°C for 20 s, annealing at 56°C for 20 s and extension at 72°C for 25 s. (b) Optimization of annealing temperature for the modified PR-PCR using a mixture of 1 U of Taq DNA polymerase and 0.1 U of KOD FX DNA polymerase. As the optimal discrimination efficiency was obtained when the annealing temperature was within a scope of 61.8°C–62.6°C, the results from 50°C to 61.2°C were not shown. The temperature 62.2°C was selected for subsequent experiments. (c) Optimization of amount of high-fidelity DNA polymerase for the modified PR-PCR with an input of 1 U of Taq DNA polymerase at an annealing temperature of 62.2°C.

    Journal: PLoS ONE

    Article Title: Modified Proofreading PCR for Detection of Point Mutations, Insertions and Deletions Using a ddNTP-Blocked Primer

    doi: 10.1371/journal.pone.0123468

    Figure Lengend Snippet: Optimization of annealing temperature and amount of high-fidelity DNA polymerase in the modified PR-PCR. (a) Discrimination efficiency of the modified PR-PCR for known mutations using a mixture of 1 U of Taq DNA polymerase and various amount (0.05, 0.1 and 0.3 U) of KOD FX DNA polymerase in the reaction. The PCR amplifications were performed in a total volume of 20 μL under cycling conditions consisting of pre-denaturation at 94°C for 2 min, followed by 30 cycles of denaturation at 94°C for 20 s, annealing at 56°C for 20 s and extension at 72°C for 25 s. (b) Optimization of annealing temperature for the modified PR-PCR using a mixture of 1 U of Taq DNA polymerase and 0.1 U of KOD FX DNA polymerase. As the optimal discrimination efficiency was obtained when the annealing temperature was within a scope of 61.8°C–62.6°C, the results from 50°C to 61.2°C were not shown. The temperature 62.2°C was selected for subsequent experiments. (c) Optimization of amount of high-fidelity DNA polymerase for the modified PR-PCR with an input of 1 U of Taq DNA polymerase at an annealing temperature of 62.2°C.

    Article Snippet: The results showed that the use of 0.1–0.15 U of high-fidelity DNA polymerase combined with 1 U of Taq DNA polymerase in per 20 μL reaction resulted in the best discrimination and amplification effects ( ).

    Techniques: Modification, Polymerase Chain Reaction

    Sensitivity and selectivity of the modified PR-PCR for mutation detection using a fusion-blocked primer and adaptor. (a) Diagram of the fusion-blocked primer and adaptor used in the reactions. (b) Primer and adaptor sequences. (c) Sensitivity of the modified PR-PCR. (d) Selectivity of the modified PR-PCR. All reactions were performed in a total volume of 25 μL containing a mixture of 1 μL of gDNA template at 50 ng/μL, 2.5 μL of (10×) Taq PCR buffer, 0.2 mM dNTPs, 0.15 U of PrimeSTAR HS DNA polymerase, 0.75 U of Taq DNA polymerase, 1 μL of DMSO and 0.3 μM each of the reverse primer, fusion-blocked forward primer and adaptor. The DNA templates were prepared by mixing different amounts of MCF-7 (WT) gDNA (from 0 to 50 ng) among HCC1937 (mutant type, MT) gDNA (from 50 to 0 ng) with concentrations from 0 to 100%. The reactions were performed under cycling conditions consisting of pre-denaturation at 94°C for 2 min, followed by 40 cycles of denaturation at 98°C for 10 s, annealing at 59°C for 30 s and extension at 72°C for 20 s.

    Journal: PLoS ONE

    Article Title: Modified Proofreading PCR for Detection of Point Mutations, Insertions and Deletions Using a ddNTP-Blocked Primer

    doi: 10.1371/journal.pone.0123468

    Figure Lengend Snippet: Sensitivity and selectivity of the modified PR-PCR for mutation detection using a fusion-blocked primer and adaptor. (a) Diagram of the fusion-blocked primer and adaptor used in the reactions. (b) Primer and adaptor sequences. (c) Sensitivity of the modified PR-PCR. (d) Selectivity of the modified PR-PCR. All reactions were performed in a total volume of 25 μL containing a mixture of 1 μL of gDNA template at 50 ng/μL, 2.5 μL of (10×) Taq PCR buffer, 0.2 mM dNTPs, 0.15 U of PrimeSTAR HS DNA polymerase, 0.75 U of Taq DNA polymerase, 1 μL of DMSO and 0.3 μM each of the reverse primer, fusion-blocked forward primer and adaptor. The DNA templates were prepared by mixing different amounts of MCF-7 (WT) gDNA (from 0 to 50 ng) among HCC1937 (mutant type, MT) gDNA (from 50 to 0 ng) with concentrations from 0 to 100%. The reactions were performed under cycling conditions consisting of pre-denaturation at 94°C for 2 min, followed by 40 cycles of denaturation at 98°C for 10 s, annealing at 59°C for 30 s and extension at 72°C for 20 s.

    Article Snippet: The results showed that the use of 0.1–0.15 U of high-fidelity DNA polymerase combined with 1 U of Taq DNA polymerase in per 20 μL reaction resulted in the best discrimination and amplification effects ( ).

    Techniques: Modification, Polymerase Chain Reaction, Mutagenesis

    Screening of DNA polymerases for single-nucleotide insertion. ( A ) The reaction for ImO N :NaN O base pair. ( B ) The reaction for ImN O :NaO N base pair. All reactions used the primer-template combination 5′-FITC-GTTCTGGATGGTCAGCGCAC-3′ (20mer) and 3′-CAAGACCTACCAGTCGCGTG X GAACGGGTG-5′ (30mer, X = ImO N , NaN O , ImN O or NaO N ). Lanes 1 and 14 indicate the 20-mer primer; lanes 2, 8, 15 and 21 indicate the results of KF (exo − ); lanes 3, 9 16, and 22 indicate those of Taq DNA polymerase; lanes 4, 10, 17 and 23 indicate those of Tth DNA polymerase; lanes 5, 11, 18 and 24 indicate those of Vent (exo − ) DNA polymerase; lanes 6, 12, 19 and 25 indicate those of Deep Vent (exo − ) DNA polymerase; and lanes 7, 13, 20 and 26 indicate those of KOD Dash DNA polymerase.

    Journal: Nucleic Acids Research

    Article Title: Unnatural imidazopyridopyrimidine:naphthyridine base pairs: selective incorporation and extension reaction by Deep Vent (exo− ) DNA polymerase

    doi: 10.1093/nar/gkp611

    Figure Lengend Snippet: Screening of DNA polymerases for single-nucleotide insertion. ( A ) The reaction for ImO N :NaN O base pair. ( B ) The reaction for ImN O :NaO N base pair. All reactions used the primer-template combination 5′-FITC-GTTCTGGATGGTCAGCGCAC-3′ (20mer) and 3′-CAAGACCTACCAGTCGCGTG X GAACGGGTG-5′ (30mer, X = ImO N , NaN O , ImN O or NaO N ). Lanes 1 and 14 indicate the 20-mer primer; lanes 2, 8, 15 and 21 indicate the results of KF (exo − ); lanes 3, 9 16, and 22 indicate those of Taq DNA polymerase; lanes 4, 10, 17 and 23 indicate those of Tth DNA polymerase; lanes 5, 11, 18 and 24 indicate those of Vent (exo − ) DNA polymerase; lanes 6, 12, 19 and 25 indicate those of Deep Vent (exo − ) DNA polymerase; and lanes 7, 13, 20 and 26 indicate those of KOD Dash DNA polymerase.

    Article Snippet: Taq DNA polymerase was purchased from TaKaRa.

    Techniques: