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TaKaRa premix taq dna polymerase
Premix Taq Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/premix taq dna polymerase/product/TaKaRa
Average 99 stars, based on 41 article reviews
Price from $9.99 to $1999.99
premix taq dna polymerase - by Bioz Stars, 2020-04
99/100 stars

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Countercurrent Chromatography:

Article Title: Halogen Bond Interactions of Novel HIV-1 Protease Inhibitors (PI) (GRL-001-15 and GRL-003-15) with the Flap of Protease Are Critical for Their Potent Activity against Wild-Type HIV-1 and Multi-PI-Resistant Variants
Article Snippet: The PCR mixture consisted of 1 μl proviral DNA solution, 10 μl Premix Taq polymerase (Premix Ex Taq version 2; TaKaRa Bio Inc., Shiga, Japan), and 10 pmol of each PCR primer in a total volume of 20 μl. .. The second-round PCR primers used for the PR-encoding region were KAPA-1 (5′-GCA GGG CCC CTA GGA AAA AGG GCT GTT GG-3′) and Ksma2.1 (5′-CCA TCC CGG GCT TTA ATT TTA CTG GTA C-3′) under the PCR conditions of an initial 1 min at 95°C, followed by 30 cycles of 30 s at 95°C, 30 s at 55°C, and 1 min at 72°C, with a final 7 min of extension at 72°C.

Molecular Cloning:

Article Title: Halogen Bond Interactions of Novel HIV-1 Protease Inhibitors (PI) (GRL-001-15 and GRL-003-15) with the Flap of Protease Are Critical for Their Potent Activity against Wild-Type HIV-1 and Multi-PI-Resistant Variants
Article Snippet: In brief, DNA was extracted from HIV-1-infected MT-4 cells by using DNAzol Direct (Molecular Research Center, Cincinnati, OH) and was subjected to molecular cloning, followed by nucleotide sequence determination. .. The PCR mixture consisted of 1 μl proviral DNA solution, 10 μl Premix Taq polymerase (Premix Ex Taq version 2; TaKaRa Bio Inc., Shiga, Japan), and 10 pmol of each PCR primer in a total volume of 20 μl.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Coxsackievirus B3 Infection Triggers Autophagy through 3 Pathways of Endoplasmic Reticulum Stress.
Article Snippet: Paragraph title: Semiquantitative RT-PCR ... PCR was performed using Premix Taq™ (R004A, TaKaRa) with the corresponding primer pairs: primers for XBP1 (human) are 5'-CTGGAACAGCAAGTGGTAGA-3' and 5'-CTGGATCCTTCTGGGTAGAC-3'; primers for β-actin (human) are 5'-TTAGTTGCGTTACACCCTTTCTTG-3' and 5'-TCACCTTCACCGTTCCAGTTT-3'.

Synthesized:

Article Title: Coxsackievirus B3 Infection Triggers Autophagy through 3 Pathways of Endoplasmic Reticulum Stress.
Article Snippet: First-strand complementary DNAs (cDNAs) were synthesized using PrimeScript RT Master Mix (RR036A, TaKaRa). .. PCR was performed using Premix Taq™ (R004A, TaKaRa) with the corresponding primer pairs: primers for XBP1 (human) are 5'-CTGGAACAGCAAGTGGTAGA-3' and 5'-CTGGATCCTTCTGGGTAGAC-3'; primers for β-actin (human) are 5'-TTAGTTGCGTTACACCCTTTCTTG-3' and 5'-TCACCTTCACCGTTCCAGTTT-3'.

CTG Assay:

Article Title: Halogen Bond Interactions of Novel HIV-1 Protease Inhibitors (PI) (GRL-001-15 and GRL-003-15) with the Flap of Protease Are Critical for Their Potent Activity against Wild-Type HIV-1 and Multi-PI-Resistant Variants
Article Snippet: Primers used for the first round of PCR with the entire Gag-PR-encoding regions of the HIV-1 genome were LTR-F1 (5′-GAT GCT ACA TAT AAG CAG CTG C-3′) and PR12 (5′-CTC GTG ACA AAT TTC TAC TAA TGC-3′). .. The PCR mixture consisted of 1 μl proviral DNA solution, 10 μl Premix Taq polymerase (Premix Ex Taq version 2; TaKaRa Bio Inc., Shiga, Japan), and 10 pmol of each PCR primer in a total volume of 20 μl.

Polymerase Chain Reaction:

Article Title: Halogen Bond Interactions of Novel HIV-1 Protease Inhibitors (PI) (GRL-001-15 and GRL-003-15) with the Flap of Protease Are Critical for Their Potent Activity against Wild-Type HIV-1 and Multi-PI-Resistant Variants
Article Snippet: .. The PCR mixture consisted of 1 μl proviral DNA solution, 10 μl Premix Taq polymerase (Premix Ex Taq version 2; TaKaRa Bio Inc., Shiga, Japan), and 10 pmol of each PCR primer in a total volume of 20 μl. ..

Article Title: Coxsackievirus B3 Infection Triggers Autophagy through 3 Pathways of Endoplasmic Reticulum Stress.
Article Snippet: .. PCR was performed using Premix Taq™ (R004A, TaKaRa) with the corresponding primer pairs: primers for XBP1 (human) are 5'-CTGGAACAGCAAGTGGTAGA-3' and 5'-CTGGATCCTTCTGGGTAGAC-3'; primers for β-actin (human) are 5'-TTAGTTGCGTTACACCCTTTCTTG-3' and 5'-TCACCTTCACCGTTCCAGTTT-3'. ..

Article Title: Root ethylene mediates rhizosphere microbial community reconstruction when chemically detecting cyanide produced by neighbouring plants
Article Snippet: Each sample was amplified in a 20-μl reaction system, which contained 0.5 μM forward and reverse primers, 1 × Premix Taq DNA polymerase (Takara, Kusatsu, Japan) and 20 ng DNA templates. .. After an initial denaturation at 95 °C for 3 min, the targeted region was amplified by 20 cycles of 94 °C for 30 s, 55 °C for 30 s, and 72 °C for 30 s, followed by a final extension at 72 °C for 1 min in a thermal cycler (GeneAmp PCR system 2700; Applied Biosystems, New York, USA).

Spectrophotometry:

Article Title: Root ethylene mediates rhizosphere microbial community reconstruction when chemically detecting cyanide produced by neighbouring plants
Article Snippet: The quantity and purity of DNA were examined with a Nanodrop ND-1000 spectrophotometer (NanoDrop Technologies, Delaware, USA). .. Each sample was amplified in a 20-μl reaction system, which contained 0.5 μM forward and reverse primers, 1 × Premix Taq DNA polymerase (Takara, Kusatsu, Japan) and 20 ng DNA templates.

DNA Extraction:

Article Title: Root ethylene mediates rhizosphere microbial community reconstruction when chemically detecting cyanide produced by neighbouring plants
Article Snippet: Paragraph title: Soil DNA extraction and 16S rRNA high-throughput sequencing ... Each sample was amplified in a 20-μl reaction system, which contained 0.5 μM forward and reverse primers, 1 × Premix Taq DNA polymerase (Takara, Kusatsu, Japan) and 20 ng DNA templates.

High Throughput Screening Assay:

Article Title: Root ethylene mediates rhizosphere microbial community reconstruction when chemically detecting cyanide produced by neighbouring plants
Article Snippet: Paragraph title: Soil DNA extraction and 16S rRNA high-throughput sequencing ... Each sample was amplified in a 20-μl reaction system, which contained 0.5 μM forward and reverse primers, 1 × Premix Taq DNA polymerase (Takara, Kusatsu, Japan) and 20 ng DNA templates.

Incubation:

Article Title: Root ethylene mediates rhizosphere microbial community reconstruction when chemically detecting cyanide produced by neighbouring plants
Article Snippet: Soil DNA extraction and 16S rRNA high-throughput sequencing Soil samples (1.0 g) from field and incubation experiments were extracted using the FastDNA SPIN Kit (MP Biomedical, California, USA) according to the manufacturer’s instructions. .. Each sample was amplified in a 20-μl reaction system, which contained 0.5 μM forward and reverse primers, 1 × Premix Taq DNA polymerase (Takara, Kusatsu, Japan) and 20 ng DNA templates.

Construct:

Article Title: Root ethylene mediates rhizosphere microbial community reconstruction when chemically detecting cyanide produced by neighbouring plants
Article Snippet: Each sample was amplified in a 20-μl reaction system, which contained 0.5 μM forward and reverse primers, 1 × Premix Taq DNA polymerase (Takara, Kusatsu, Japan) and 20 ng DNA templates. .. Amplicon sequencing libraries were constructed using the MiSeq Reagent Kit v3 according to the manufacturer’s instructions.

Sequencing:

Article Title: Halogen Bond Interactions of Novel HIV-1 Protease Inhibitors (PI) (GRL-001-15 and GRL-003-15) with the Flap of Protease Are Critical for Their Potent Activity against Wild-Type HIV-1 and Multi-PI-Resistant Variants
Article Snippet: In brief, DNA was extracted from HIV-1-infected MT-4 cells by using DNAzol Direct (Molecular Research Center, Cincinnati, OH) and was subjected to molecular cloning, followed by nucleotide sequence determination. .. The PCR mixture consisted of 1 μl proviral DNA solution, 10 μl Premix Taq polymerase (Premix Ex Taq version 2; TaKaRa Bio Inc., Shiga, Japan), and 10 pmol of each PCR primer in a total volume of 20 μl.

Article Title: Root ethylene mediates rhizosphere microbial community reconstruction when chemically detecting cyanide produced by neighbouring plants
Article Snippet: Paragraph title: Soil DNA extraction and 16S rRNA high-throughput sequencing ... Each sample was amplified in a 20-μl reaction system, which contained 0.5 μM forward and reverse primers, 1 × Premix Taq DNA polymerase (Takara, Kusatsu, Japan) and 20 ng DNA templates.

Amplification:

Article Title: Root ethylene mediates rhizosphere microbial community reconstruction when chemically detecting cyanide produced by neighbouring plants
Article Snippet: .. Each sample was amplified in a 20-μl reaction system, which contained 0.5 μM forward and reverse primers, 1 × Premix Taq DNA polymerase (Takara, Kusatsu, Japan) and 20 ng DNA templates. .. After an initial denaturation at 95 °C for 3 min, the targeted region was amplified by 20 cycles of 94 °C for 30 s, 55 °C for 30 s, and 72 °C for 30 s, followed by a final extension at 72 °C for 1 min in a thermal cycler (GeneAmp PCR system 2700; Applied Biosystems, New York, USA).

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  • 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, 2020-04
    99/100 stars
      Buy from Supplier

    99
    TaKaRa premix taq
    <t>RT-PCR</t> results for the expression of CIC-DUX4 in the tumor. A) The initial PCR with Premix <t>Taq</t> and the primer set CIC-4105F/DUX4-1538R (lane 1) as well as the nested PCR with the primers CIC-4283F/DUX4-1507R (lane 2) did not amplify any cDNA fragments. The primer set CIC-4238F/CIC-4958R (lane 3) amplified a CIC cDNA fragment suggesting the good quality of the synthesized cDNA. Lane 4, Blank, no RNA in cDNA synthesis. B) PCR amplifications using the PrimeSTAR GXL DNA polymerase and the primer combinations CIC-4377F/DUX4-1151R (lane 5) and CIC-4453F/DUX4-1053R (lane 6). Lane 7, Blank, no RNA in cDNA synthesis. M, 1 Kb DNA ladder (GeneRuler, Fermentas). C) Partial sequence chromatogram of the amplified cDNA fragment showing that CIC is fused to DUX4.
    Premix Taq, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 245 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/premix taq/product/TaKaRa
    Average 99 stars, based on 245 article reviews
    Price from $9.99 to $1999.99
    premix taq - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

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    (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:

    RT-PCR results for the expression of CIC-DUX4 in the tumor. A) The initial PCR with Premix Taq and the primer set CIC-4105F/DUX4-1538R (lane 1) as well as the nested PCR with the primers CIC-4283F/DUX4-1507R (lane 2) did not amplify any cDNA fragments. The primer set CIC-4238F/CIC-4958R (lane 3) amplified a CIC cDNA fragment suggesting the good quality of the synthesized cDNA. Lane 4, Blank, no RNA in cDNA synthesis. B) PCR amplifications using the PrimeSTAR GXL DNA polymerase and the primer combinations CIC-4377F/DUX4-1151R (lane 5) and CIC-4453F/DUX4-1053R (lane 6). Lane 7, Blank, no RNA in cDNA synthesis. M, 1 Kb DNA ladder (GeneRuler, Fermentas). C) Partial sequence chromatogram of the amplified cDNA fragment showing that CIC is fused to DUX4.

    Journal: PLoS ONE

    Article Title: The "Grep" Command But Not FusionMap, FusionFinder or ChimeraScan Captures the CIC-DUX4 Fusion Gene from Whole Transcriptome Sequencing Data on a Small Round Cell Tumor with t(4;19)(q35;q13)

    doi: 10.1371/journal.pone.0099439

    Figure Lengend Snippet: RT-PCR results for the expression of CIC-DUX4 in the tumor. A) The initial PCR with Premix Taq and the primer set CIC-4105F/DUX4-1538R (lane 1) as well as the nested PCR with the primers CIC-4283F/DUX4-1507R (lane 2) did not amplify any cDNA fragments. The primer set CIC-4238F/CIC-4958R (lane 3) amplified a CIC cDNA fragment suggesting the good quality of the synthesized cDNA. Lane 4, Blank, no RNA in cDNA synthesis. B) PCR amplifications using the PrimeSTAR GXL DNA polymerase and the primer combinations CIC-4377F/DUX4-1151R (lane 5) and CIC-4453F/DUX4-1053R (lane 6). Lane 7, Blank, no RNA in cDNA synthesis. M, 1 Kb DNA ladder (GeneRuler, Fermentas). C) Partial sequence chromatogram of the amplified cDNA fragment showing that CIC is fused to DUX4.

    Article Snippet: Initially, the 25 µL PCR-volume contained 12.5 µL of Premix Taq (Takara Bio Europe/SAS, Saint-Germain-en-Laye, France), 1 µL of the synthesized cDNA, and 0.4 µM of each of the forward CIC-4105F and reverse DUX4-1538R primers.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Polymerase Chain Reaction, Nested PCR, Amplification, Synthesized, Sequencing