ex taq dna polymerase  (TaKaRa)

 
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    Name:
    Premix Ex Taq DNA Polymerase
    Description:
    Premix Ex Taq DNA Polymerase Perfect Real Time is a 2X RT PCR kit specifically designed for fast and sensitive real time PCR via either intercalating green dye Real Time PCR qPCR or probe based qPCR assays TB Green dye and probes for probe 5 nuclease based assays are not included in this RT PCR kit The Premix Ex Taq DNA Polymerase Perfect Real Time kit consists of our high fidelity and high performance Takara Ex Taq Hot Start polymerase and a real time buffer that ensures superior specificity and increased amplification efficiency during RT PCR qPCR Antibody mediated hot start technology prevents nonspecific amplification due to mispriming and or the formation of primer dimers during room temperature reaction assembly Once the Taq antibody polymerase complex is denatured during the first cycling step the Premix Ex Taq polymerase can initiate DNA synthesis
    Catalog Number:
    rr039b
    Price:
    None
    Size:
    400 Rxns
    Category:
    Premix Ex Taq DNA Polymerase Perfect Real Time qPCR with probe detection Real time PCR kits Real time PCR
    Buy from Supplier
    Name:
    TaKaRa Ex Taq DNA Polymerase
    Description:
    TaKaRa Ex Taq DNA Polymerase combines the proven performance of Takara Taq polymerase with the proofreading activity of an efficient 3 to 5 exonuclease for high sensitivity high efficiency PCR reactions It can also be used for long range PCR up to 20 kb from genomic DNA templates and up to 30 kb from lambda DNA templates Ex Taq polymerase has a higher fidelity than standard Taq with a mutation rate approximately 4 5 times lower as determined by the Kunkel method Ex Taq polymerase is supplied with optimized 10X buffer with or without Mg2 and dNTPs
    Catalog Number:
    rr001c
    Price:
    None
    Size:
    3 000 Units
    Category:
    Ex Taq polymerase Ex Taq products High yield PCR PCR
    Buy from Supplier


    Structured Review

    TaKaRa ex taq dna polymerase
    TaKaRa Ex Taq DNA Polymerase combines the proven performance of Takara Taq polymerase with the proofreading activity of an efficient 3 to 5 exonuclease for high sensitivity high efficiency PCR reactions It can also be used for long range PCR up to 20 kb from genomic DNA templates and up to 30 kb from lambda DNA templates Ex Taq polymerase has a higher fidelity than standard Taq with a mutation rate approximately 4 5 times lower as determined by the Kunkel method Ex Taq polymerase is supplied with optimized 10X buffer with or without Mg2 and dNTPs
    https://www.bioz.com/result/ex taq dna polymerase/product/TaKaRa
    Average 99 stars, based on 840 article reviews
    Price from $9.99 to $1999.99
    ex taq dna polymerase - by Bioz Stars, 2020-01
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli
    Article Snippet: .. DNA cloning procedure All DNA cloning experiments were carried out according to the standard procedures , and Ex-Taq polymerase and Pfu DNA polymerase (Takara, Japan) were successfully used for the polymerase chain reactions (PCR). .. The pCzFv, pCzFvHAP and pCzFvLAP vectors were constructed previously as shown in (IG Therapy, South Korea. unpublished).

    Centrifugation:

    Article Title: Multiple primer extension by DNA polymerase on a novel plastic DNA array coated with a biocompatible polymer
    Article Snippet: Primer extension using the synthetic oligonucleotide template DNA amplification was initiated on the PrimeSurface® plastic slides with a primer extension reaction mixture containing 1 U/100 μl of EX Taq polymerase in 1× EX Taq buffer (TaKaRa Biosciences Co. Ltd, Otsu, Japan), 0.05 mM each of dATP, dCTP, dGTP (GibcoBRL® ) and 0.05 mM Cy3-labeled dUTP, supplemented with 5′-Cy5-labeled target DNA mixture as a template from 0.1 to 1000 pM. .. DNA arrays were washed with pre-prepared washing buffer as described previously and dried by centrifugation for 2 min at 200 g .

    Amplification:

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: .. For fluorescent genotyping (GeneScan) analysis, the nucleotide 1150–1358 fragment of the WASP genomic DNA was amplified from 300 ng of DNA extracted from PBMCs, immunomagnetic bead-purified CD3+ T lymphocytes, CD3+ cell–depleted PMNs and sorted CD20+ B lymphocytes, CD56+ NK cells, and CD14+ monocytes, using 20 pmol each of the 5′-TCC AGC TAC TGG ACG TTC TG-3′ forward primer and the FAM-labeled 5′-TTC CCT GCC GGA TTT GAT-3′ reverse primer in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq Buffer, 2 mM Mg2+ , and 200 μM dNTPs (all from Takara Bio Inc., Shiga, Japan). .. One microliter of the amplification product was then combined to 12 μl of deionized formamide and 0.5 μl of GeneScan size standard (Perkin-Elmer Applied Biosystems) and analyzed on a ABI Prism 310 Genetic Analyzer using the GeneScan Analysis Software (Perkin-Elmer Applied Biosystems).

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: .. Briefly, 50 ng of DNA extracted from sorted WASP+ and WASP– T lymphocytes was subjected to PCR amplification using a TCR Vβ consensus primer [Vβ pan: 5′-CTC GAA TTC T(T/G) T(A/T) (C/T)T GGT A(C/T) C(G/A)(T/A)CA-3′; 30 pmol] and a TCR-Jβ family-specific primer (Jβ 2.1: 5′-ACG-GTG-AGC-CGT-GTC-CC-3′; 10 pmol) ( ) in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq buffer, 2 mM Mg2+ , and 200 μM dNTP (all from Takara Bio Inc.). .. One microliter of the amplification product was then subjected to a seminested reaction using the same amplification conditions, but using a FAM-labeled Vβ pan primer in combination with a nested Jβ 2.1–specifc primer (5′-TGA-GCC-GTG-TCC-CTG-GCC-CGA-A-3′) ( ).

    Article Title: Multiple primer extension by DNA polymerase on a novel plastic DNA array coated with a biocompatible polymer
    Article Snippet: .. Primer extension using the synthetic oligonucleotide template DNA amplification was initiated on the PrimeSurface® plastic slides with a primer extension reaction mixture containing 1 U/100 μl of EX Taq polymerase in 1× EX Taq buffer (TaKaRa Biosciences Co. Ltd, Otsu, Japan), 0.05 mM each of dATP, dCTP, dGTP (GibcoBRL® ) and 0.05 mM Cy3-labeled dUTP, supplemented with 5′-Cy5-labeled target DNA mixture as a template from 0.1 to 1000 pM. .. Our homemade hybridization cassette was immersed in 50 μl of the reaction mixture in a frame seal chamber.

    Article Title: Molecular Cloning and Characterization of Taurocyamine Kinase from Clonorchis sinensis: A Candidate Chemotherapeutic Target
    Article Snippet: .. Amplification of 3′-cDNA end of C. sinensis PK D2 domain Polymerase chain reaction (PCR) was carried out with reaction mixture containing cDNA, 10 pmol of each primer, 2 µl of 2.5 mM of dNTPs, 1 U of Ex Taq polymerase, 2.5 µl of 10× Ex Taq buffer (TaKaRa, Tokyo, Japan). .. Thermal cycles were prepared as follows: initial denaturation at 94°C for 2 min, followed by 35 cycles of 94°C for 30 s, annealing at 50°C for 35 s, and extension at 72°C for 2 min, and a final extension at 72°C for 4 min. PCR was done in a thermal cycler, MyCycler (BioRad, Foster, USA).

    Article Title: Fluvoxamine alleviates ER stress via induction of Sigma-1 receptor
    Article Snippet: PCR was performed in 50 μ l containing 0.8 μ M of each primer, 0.2 mM dNTPs, 1.25 U of Ex-Taq polymerase, and 10 × PCR buffer (Takara). .. The primer sets used for amplification were as follows: Sig-1R forward, 5′-GCCTTCTCTCGTCTGATCGT-3′ and Sig-1R reverse, 5′-GCCAAAGAGGTAGGTGGTGA-3′ β -actin forward, 5′-GTTTGAGACCTCAACACC-3′ and β -actin reverse, 5′-GTGGTGGTGAAGCTTAG-3′, X-box binding protein 1 (XBP-1), 5′-TAAGACAGCGCTTGGGGATGG-3′, and 5′-CAGAATCCATGGGGAGATGTT-3′.

    Article Title: Agrobacterium tumefaciens – Mediated transformation of Woodfordia fruticosa (L.) Kurz
    Article Snippet: The 90 bp fragment specific to uidA gene and 1 kb hpt gene fragment were amplified using the following primer pairs: uidA : Forward-5′-CGACGGCCTGTGGGCATTTCA-3′ and Reverse-5′-TGGTCGTGCACCATCAGCAC-3′; hpt : Forward-5′-TAGAAAAAGCCTGAACTCACCG-3′ and Reverse-5′-TATTTCTTTGCCCTCGGACG-3′. .. PCR reactions were carried out in 20 μl reaction mixture containing 0.5 units of Ex Taq polymerase and 1× Taq buffer (Takara, Dalian, China), 0.2 mM of each dNTP, 0.5 μM of each primer, and 50 ng of template DNA.

    Article Title: Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli
    Article Snippet: DNA cloning procedure All DNA cloning experiments were carried out according to the standard procedures , and Ex-Taq polymerase and Pfu DNA polymerase (Takara, Japan) were successfully used for the polymerase chain reactions (PCR). .. The VH and the VLκ chain genes were PCR amplified from SP112 Fab clone ( ) at the condition of 35 cycles of 94℃ 1 min, 55℃ 1 min, 72℃ 1 min, followed by 72℃ soaking for 10 min using human Ig-specific VH and JH , or VLκ and Jκ primers synthesized according to the previous report with slight modification (VH sense: 5'-GGG GGCCCAGCCGGCC ATGGCCGAGGTGCAGCTGGTGGAGTCTGG-3', JH antisense: 5'-GGG GGCCACATTGGCC GATGAGGAGACGGTGACCAKGGTBCCTTGGCCCCA-3', Vκ sense: VLκ forward: 5'-GGG GTCGAC ATGGAAATTGAGTTGACGCAGTCTCC-3', Jκ antisense: 5'-GGG CCGCGG ATACGTTTGATHTCCASYTTGGTCCC-3'; where Sfi I, Sal I and Sac II recognition sites were underlined, and degeneracy is denoted as follows: H=A, C or T; S=G or C; Y=C or T; K=G or T; B=G, T or C) ( ).

    Article Title: A Novel A1088T Mutation in the Glucose-6-Phosphate Dehydrogenase Gene Detected by RT-PCR Combined with DNA Sequencing
    Article Snippet: .. The PCR amplification was performed with EX-Taq polymerase and GC buffer(Takara) in a thermocycler(Biometra, Germany) for an initial denaturing cycle at 94 °C for 2 min, followed by 30 cycles (94 °C for 30 s, 59 °C for 30 s, 72 °C for 2 min), and a final extension step at 72 °C for 8 min. Genomic DNA was used to verify the novel mutation site and analyze mutations in the promoter region (from −509 bp to +26 bp). .. The promoter region of human G6PD DNA was amplified by PCR with primer pairs 5′-CCACGGATGGAACCCTGTC-3′/5′-CCGAAGTGTACGACCGTTTC-3′.

    Article Title: High Prevalence of Plasmid-Mediated Quinolone Resistance Determinants qnr, aac(6?)-Ib-cr, and qepA among Ceftiofur-Resistant Enterobacteriaceae Isolates from Companion and Food-Producing Animals ▿
    Article Snippet: .. The PCR amplifications were performed in 25-μl volumes containing 2.5 mM MgCl2 , 0.5 U of Ex Taq polymerase, 0.25 mM each deoxynucleoside triphosphate, 2.5 μl of 10× amplification buffer, 10 ng of crude template DNA, and 25 pmol each of two opposing primers, primers ERIC1R (5′-ATGTAAGCTCCTGGGGATTCA-3′) and ERIC2 (5′-AAGTAAGTGACTGGGGTGAGCG-3′) (TaKaRa Bio, Dalian, China) ( ). .. A negative control consisting of the same reaction mixture without a DNA template was included in each PCR.

    Article Title: High-Throughput Discovery of Chloroplast and Mitochondrial DNA Polymorphisms in Brassicaceae Species by ORG-EcoTILLING
    Article Snippet: .. The first PCR amplification was performed in a 15 µL reaction volume using 60 ng DNA template, 1.5 µL 10× Ex-buffer, 0.2 mM dNTPs, 0.75 U Ex-Taq polymerase and 0.25 µL 10 µM each primer [Ex-Taq polymerase and Ex-buffer both purchased from TAKARA Biotechnology (Dalian) CO., LTD.]. ..

    Positive Control:

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: Briefly, 50 ng of DNA extracted from sorted WASP+ and WASP– T lymphocytes was subjected to PCR amplification using a TCR Vβ consensus primer [Vβ pan: 5′-CTC GAA TTC T(T/G) T(A/T) (C/T)T GGT A(C/T) C(G/A)(T/A)CA-3′; 30 pmol] and a TCR-Jβ family-specific primer (Jβ 2.1: 5′-ACG-GTG-AGC-CGT-GTC-CC-3′; 10 pmol) ( ) in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq buffer, 2 mM Mg2+ , and 200 μM dNTP (all from Takara Bio Inc.). .. DNA extracted from MOLT-4 cells with known clonal TCR-Jβ 2.1 rearrangement was used as positive control.

    Synthesized:

    Article Title: Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli
    Article Snippet: DNA cloning procedure All DNA cloning experiments were carried out according to the standard procedures , and Ex-Taq polymerase and Pfu DNA polymerase (Takara, Japan) were successfully used for the polymerase chain reactions (PCR). .. The VH and the VLκ chain genes were PCR amplified from SP112 Fab clone ( ) at the condition of 35 cycles of 94℃ 1 min, 55℃ 1 min, 72℃ 1 min, followed by 72℃ soaking for 10 min using human Ig-specific VH and JH , or VLκ and Jκ primers synthesized according to the previous report with slight modification (VH sense: 5'-GGG GGCCCAGCCGGCC ATGGCCGAGGTGCAGCTGGTGGAGTCTGG-3', JH antisense: 5'-GGG GGCCACATTGGCC GATGAGGAGACGGTGACCAKGGTBCCTTGGCCCCA-3', Vκ sense: VLκ forward: 5'-GGG GTCGAC ATGGAAATTGAGTTGACGCAGTCTCC-3', Jκ antisense: 5'-GGG CCGCGG ATACGTTTGATHTCCASYTTGGTCCC-3'; where Sfi I, Sal I and Sac II recognition sites were underlined, and degeneracy is denoted as follows: H=A, C or T; S=G or C; Y=C or T; K=G or T; B=G, T or C) ( ).

    Construct:

    Article Title: Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli
    Article Snippet: DNA cloning procedure All DNA cloning experiments were carried out according to the standard procedures , and Ex-Taq polymerase and Pfu DNA polymerase (Takara, Japan) were successfully used for the polymerase chain reactions (PCR). .. The pCzFv, pCzFvHAP and pCzFvLAP vectors were constructed previously as shown in (IG Therapy, South Korea. unpublished).

    Expressing:

    Article Title: Cloning and expression of visfatin and screening of oligopeptides binding with visfatin
    Article Snippet: Prokaryotic expression vector pQE-30Xa and E.coli strains M15 [pREP4] were purchased from Qiagen. .. The enzymes Sal I, Stu I, Pvu II, Ex Taq polymerase and dNTP were purchased from Takara Bio Inc. M-MLV reverse transcriptase and T4 DNA ligase were purchased from Promega.

    Article Title: Prostaglandin E2-Induced COX-2 Expressions via EP2 and EP4 Signaling Pathways in Human LoVo Colon Cancer Cells
    Article Snippet: .. The Expression of EP1–EP4 in LoVo Colon Cancer Cells Was Detected by Reverse Transcription PCR (RT-PCR) RT-PCR was carried out with the Ex Taq polymerase using Ex Taq Master Mix kit (Clontech Laboratries, Mountain View, CA, USA). ..

    Article Title: Display of Bombyx mori Alcohol Dehydrogenases on the Bacillus subtilis Spore Surface to Enhance Enzymatic Activity under Adverse Conditions
    Article Snippet: The expression vector pET-30a(+) and Escherichia coli strains DH5α and BL21(DE3) were obtained from Novagen (CA, USA). .. Ex Taq polymerase, restriction enzymes, T4 DNA ligase and the subcloning vector pMD18-T were purchased from TaKaRa (Dalian, China).

    Article Title: Agrobacterium tumefaciens – Mediated transformation of Woodfordia fruticosa (L.) Kurz
    Article Snippet: PCR reactions were carried out in 20 μl reaction mixture containing 0.5 units of Ex Taq polymerase and 1× Taq buffer (Takara, Dalian, China), 0.2 mM of each dNTP, 0.5 μM of each primer, and 50 ng of template DNA. .. The PCR conditions for hpt gene detection were set as initial-denaturation at 94 °C for 5 min, 30 cycles of denaturation at 95 °C for 40 s, annealing at 56 °C for 1 min, extension at 72 °C for 40 s and final extension at 72 °C for 15 min. To know the expression of transgenes by RT-PCR (reverse transcription-PCR), total RNA was isolated from in vitro regenerated transgenic plant lines following the established standard protocol and treated with DNase I (Takara, Dalian, China) to remove DNA traces.

    Modification:

    Article Title: Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli
    Article Snippet: DNA cloning procedure All DNA cloning experiments were carried out according to the standard procedures , and Ex-Taq polymerase and Pfu DNA polymerase (Takara, Japan) were successfully used for the polymerase chain reactions (PCR). .. The VH and the VLκ chain genes were PCR amplified from SP112 Fab clone ( ) at the condition of 35 cycles of 94℃ 1 min, 55℃ 1 min, 72℃ 1 min, followed by 72℃ soaking for 10 min using human Ig-specific VH and JH , or VLκ and Jκ primers synthesized according to the previous report with slight modification (VH sense: 5'-GGG GGCCCAGCCGGCC ATGGCCGAGGTGCAGCTGGTGGAGTCTGG-3', JH antisense: 5'-GGG GGCCACATTGGCC GATGAGGAGACGGTGACCAKGGTBCCTTGGCCCCA-3', Vκ sense: VLκ forward: 5'-GGG GTCGAC ATGGAAATTGAGTTGACGCAGTCTCC-3', Jκ antisense: 5'-GGG CCGCGG ATACGTTTGATHTCCASYTTGGTCCC-3'; where Sfi I, Sal I and Sac II recognition sites were underlined, and degeneracy is denoted as follows: H=A, C or T; S=G or C; Y=C or T; K=G or T; B=G, T or C) ( ).

    Transformation Assay:

    Article Title: Display of Bombyx mori Alcohol Dehydrogenases on the Bacillus subtilis Spore Surface to Enhance Enzymatic Activity under Adverse Conditions
    Article Snippet: Ex Taq polymerase, restriction enzymes, T4 DNA ligase and the subcloning vector pMD18-T were purchased from TaKaRa (Dalian, China). .. Preparation and transformation of B. subtilis strain 168 (trp-) competent cells were performed as previously described .

    Article Title: Agrobacterium tumefaciens – Mediated transformation of Woodfordia fruticosa (L.) Kurz
    Article Snippet: Paragraph title: Molecular confirmation of putatively transformed plants ... PCR reactions were carried out in 20 μl reaction mixture containing 0.5 units of Ex Taq polymerase and 1× Taq buffer (Takara, Dalian, China), 0.2 mM of each dNTP, 0.5 μM of each primer, and 50 ng of template DNA.

    Hybridization:

    Article Title: Multiple primer extension by DNA polymerase on a novel plastic DNA array coated with a biocompatible polymer
    Article Snippet: Primer extension using the synthetic oligonucleotide template DNA amplification was initiated on the PrimeSurface® plastic slides with a primer extension reaction mixture containing 1 U/100 μl of EX Taq polymerase in 1× EX Taq buffer (TaKaRa Biosciences Co. Ltd, Otsu, Japan), 0.05 mM each of dATP, dCTP, dGTP (GibcoBRL® ) and 0.05 mM Cy3-labeled dUTP, supplemented with 5′-Cy5-labeled target DNA mixture as a template from 0.1 to 1000 pM. .. Our homemade hybridization cassette was immersed in 50 μl of the reaction mixture in a frame seal chamber.

    Generated:

    Article Title: A Novel A1088T Mutation in the Glucose-6-Phosphate Dehydrogenase Gene Detected by RT-PCR Combined with DNA Sequencing
    Article Snippet: The cDNA was generated with 1 µg of total RNA, random hexadeoxynucleotide primer and RAV-2 reverse transcriptase (Takara, Dalian, China). .. The PCR amplification was performed with EX-Taq polymerase and GC buffer(Takara) in a thermocycler(Biometra, Germany) for an initial denaturing cycle at 94 °C for 2 min, followed by 30 cycles (94 °C for 30 s, 59 °C for 30 s, 72 °C for 2 min), and a final extension step at 72 °C for 8 min. Genomic DNA was used to verify the novel mutation site and analyze mutations in the promoter region (from −509 bp to +26 bp).

    Polymerase Chain Reaction:

    Article Title: Immune responses of Helicoverpa armigera to different kinds of pathogens
    Article Snippet: .. Chemicals The chemicals were obtained from the following separate companies: Unizol reagent (Biostar Company, Shanghai, China); Reverse Transcriptase (RT) Kit (Promega Biosciences, Madison WI, USA); PCR purification kit (Shengong, Shanghai, China); Ex Taq Polymerase and SYBR® Premix EX Taq (TaKaRa Biotech, Dalian, China); and 4'-6-Diamidino-2-phenylindole dihydrochloride (DAPI, 1 μg/ml in water, San Jose, United States). ..

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: Sequencing was performed on gel-purified PCR products or PCR products subcloned in the pCR2.1 vector (Invitrogen Corp., Carlsbad, California, USA) using the ABI Prism BigDye Terminator Cycle sequencing kit on an ABI Prism 310 Genetic Analyzer (Perkin-Elmer Applied Biosystems, Foster City, California, USA), as described previously ( ). .. For fluorescent genotyping (GeneScan) analysis, the nucleotide 1150–1358 fragment of the WASP genomic DNA was amplified from 300 ng of DNA extracted from PBMCs, immunomagnetic bead-purified CD3+ T lymphocytes, CD3+ cell–depleted PMNs and sorted CD20+ B lymphocytes, CD56+ NK cells, and CD14+ monocytes, using 20 pmol each of the 5′-TCC AGC TAC TGG ACG TTC TG-3′ forward primer and the FAM-labeled 5′-TTC CCT GCC GGA TTT GAT-3′ reverse primer in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq Buffer, 2 mM Mg2+ , and 200 μM dNTPs (all from Takara Bio Inc., Shiga, Japan).

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: .. Briefly, 50 ng of DNA extracted from sorted WASP+ and WASP– T lymphocytes was subjected to PCR amplification using a TCR Vβ consensus primer [Vβ pan: 5′-CTC GAA TTC T(T/G) T(A/T) (C/T)T GGT A(C/T) C(G/A)(T/A)CA-3′; 30 pmol] and a TCR-Jβ family-specific primer (Jβ 2.1: 5′-ACG-GTG-AGC-CGT-GTC-CC-3′; 10 pmol) ( ) in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq buffer, 2 mM Mg2+ , and 200 μM dNTP (all from Takara Bio Inc.). .. One microliter of the amplification product was then subjected to a seminested reaction using the same amplification conditions, but using a FAM-labeled Vβ pan primer in combination with a nested Jβ 2.1–specifc primer (5′-TGA-GCC-GTG-TCC-CTG-GCC-CGA-A-3′) ( ).

    Article Title: Multiple primer extension by DNA polymerase on a novel plastic DNA array coated with a biocompatible polymer
    Article Snippet: Primer extension using the synthetic oligonucleotide template DNA amplification was initiated on the PrimeSurface® plastic slides with a primer extension reaction mixture containing 1 U/100 μl of EX Taq polymerase in 1× EX Taq buffer (TaKaRa Biosciences Co. Ltd, Otsu, Japan), 0.05 mM each of dATP, dCTP, dGTP (GibcoBRL® ) and 0.05 mM Cy3-labeled dUTP, supplemented with 5′-Cy5-labeled target DNA mixture as a template from 0.1 to 1000 pM. .. Thermocycling was carried out as follows: 95°C for 5 min and 30 cycles (a denaturing step at 95°C for 1 min, an annealing step at 50°C for 3 min) using a GeneAmp® PCR System 9700 (Applied Biosystems).

    Article Title: Molecular Cloning and Characterization of Taurocyamine Kinase from Clonorchis sinensis: A Candidate Chemotherapeutic Target
    Article Snippet: .. Amplification of 3′-cDNA end of C. sinensis PK D2 domain Polymerase chain reaction (PCR) was carried out with reaction mixture containing cDNA, 10 pmol of each primer, 2 µl of 2.5 mM of dNTPs, 1 U of Ex Taq polymerase, 2.5 µl of 10× Ex Taq buffer (TaKaRa, Tokyo, Japan). .. Thermal cycles were prepared as follows: initial denaturation at 94°C for 2 min, followed by 35 cycles of 94°C for 30 s, annealing at 50°C for 35 s, and extension at 72°C for 2 min, and a final extension at 72°C for 4 min. PCR was done in a thermal cycler, MyCycler (BioRad, Foster, USA).

    Article Title: Prostaglandin E2-Induced COX-2 Expressions via EP2 and EP4 Signaling Pathways in Human LoVo Colon Cancer Cells
    Article Snippet: .. The Expression of EP1–EP4 in LoVo Colon Cancer Cells Was Detected by Reverse Transcription PCR (RT-PCR) RT-PCR was carried out with the Ex Taq polymerase using Ex Taq Master Mix kit (Clontech Laboratries, Mountain View, CA, USA). ..

    Article Title: Fluvoxamine alleviates ER stress via induction of Sigma-1 receptor
    Article Snippet: .. PCR was performed in 50 μ l containing 0.8 μ M of each primer, 0.2 mM dNTPs, 1.25 U of Ex-Taq polymerase, and 10 × PCR buffer (Takara). .. The primer sets used for amplification were as follows: Sig-1R forward, 5′-GCCTTCTCTCGTCTGATCGT-3′ and Sig-1R reverse, 5′-GCCAAAGAGGTAGGTGGTGA-3′ β -actin forward, 5′-GTTTGAGACCTCAACACC-3′ and β -actin reverse, 5′-GTGGTGGTGAAGCTTAG-3′, X-box binding protein 1 (XBP-1), 5′-TAAGACAGCGCTTGGGGATGG-3′, and 5′-CAGAATCCATGGGGAGATGTT-3′.

    Article Title: Agrobacterium tumefaciens – Mediated transformation of Woodfordia fruticosa (L.) Kurz
    Article Snippet: .. PCR reactions were carried out in 20 μl reaction mixture containing 0.5 units of Ex Taq polymerase and 1× Taq buffer (Takara, Dalian, China), 0.2 mM of each dNTP, 0.5 μM of each primer, and 50 ng of template DNA. .. The PCR cycling conditions for uidA included initial-denaturation at 95 °C for 5 min, 30 cycles of denaturation at 94 °C for 40 s, annealing at 58 °C for 40 s, extension at 72 °C for 1 min and final extension at 72 °C for 10 min.

    Article Title: Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli
    Article Snippet: .. DNA cloning procedure All DNA cloning experiments were carried out according to the standard procedures , and Ex-Taq polymerase and Pfu DNA polymerase (Takara, Japan) were successfully used for the polymerase chain reactions (PCR). .. The pCzFv, pCzFvHAP and pCzFvLAP vectors were constructed previously as shown in (IG Therapy, South Korea. unpublished).

    Article Title: A Novel A1088T Mutation in the Glucose-6-Phosphate Dehydrogenase Gene Detected by RT-PCR Combined with DNA Sequencing
    Article Snippet: .. The PCR amplification was performed with EX-Taq polymerase and GC buffer(Takara) in a thermocycler(Biometra, Germany) for an initial denaturing cycle at 94 °C for 2 min, followed by 30 cycles (94 °C for 30 s, 59 °C for 30 s, 72 °C for 2 min), and a final extension step at 72 °C for 8 min. Genomic DNA was used to verify the novel mutation site and analyze mutations in the promoter region (from −509 bp to +26 bp). .. The promoter region of human G6PD DNA was amplified by PCR with primer pairs 5′-CCACGGATGGAACCCTGTC-3′/5′-CCGAAGTGTACGACCGTTTC-3′.

    Article Title: High Prevalence of Plasmid-Mediated Quinolone Resistance Determinants qnr, aac(6?)-Ib-cr, and qepA among Ceftiofur-Resistant Enterobacteriaceae Isolates from Companion and Food-Producing Animals ▿
    Article Snippet: .. The PCR amplifications were performed in 25-μl volumes containing 2.5 mM MgCl2 , 0.5 U of Ex Taq polymerase, 0.25 mM each deoxynucleoside triphosphate, 2.5 μl of 10× amplification buffer, 10 ng of crude template DNA, and 25 pmol each of two opposing primers, primers ERIC1R (5′-ATGTAAGCTCCTGGGGATTCA-3′) and ERIC2 (5′-AAGTAAGTGACTGGGGTGAGCG-3′) (TaKaRa Bio, Dalian, China) ( ). .. A negative control consisting of the same reaction mixture without a DNA template was included in each PCR.

    Article Title: High-Throughput Discovery of Chloroplast and Mitochondrial DNA Polymorphisms in Brassicaceae Species by ORG-EcoTILLING
    Article Snippet: .. The first PCR amplification was performed in a 15 µL reaction volume using 60 ng DNA template, 1.5 µL 10× Ex-buffer, 0.2 mM dNTPs, 0.75 U Ex-Taq polymerase and 0.25 µL 10 µM each primer [Ex-Taq polymerase and Ex-buffer both purchased from TAKARA Biotechnology (Dalian) CO., LTD.]. ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Prostaglandin E2-Induced COX-2 Expressions via EP2 and EP4 Signaling Pathways in Human LoVo Colon Cancer Cells
    Article Snippet: .. The Expression of EP1–EP4 in LoVo Colon Cancer Cells Was Detected by Reverse Transcription PCR (RT-PCR) RT-PCR was carried out with the Ex Taq polymerase using Ex Taq Master Mix kit (Clontech Laboratries, Mountain View, CA, USA). ..

    Article Title: Fluvoxamine alleviates ER stress via induction of Sigma-1 receptor
    Article Snippet: Paragraph title: RNA extraction and semiquantitative RT-PCR ... PCR was performed in 50 μ l containing 0.8 μ M of each primer, 0.2 mM dNTPs, 1.25 U of Ex-Taq polymerase, and 10 × PCR buffer (Takara).

    Article Title: Agrobacterium tumefaciens – Mediated transformation of Woodfordia fruticosa (L.) Kurz
    Article Snippet: PCR reactions were carried out in 20 μl reaction mixture containing 0.5 units of Ex Taq polymerase and 1× Taq buffer (Takara, Dalian, China), 0.2 mM of each dNTP, 0.5 μM of each primer, and 50 ng of template DNA. .. The PCR conditions for hpt gene detection were set as initial-denaturation at 94 °C for 5 min, 30 cycles of denaturation at 95 °C for 40 s, annealing at 56 °C for 1 min, extension at 72 °C for 40 s and final extension at 72 °C for 15 min. To know the expression of transgenes by RT-PCR (reverse transcription-PCR), total RNA was isolated from in vitro regenerated transgenic plant lines following the established standard protocol and treated with DNase I (Takara, Dalian, China) to remove DNA traces.

    Binding Assay:

    Article Title: Fluvoxamine alleviates ER stress via induction of Sigma-1 receptor
    Article Snippet: PCR was performed in 50 μ l containing 0.8 μ M of each primer, 0.2 mM dNTPs, 1.25 U of Ex-Taq polymerase, and 10 × PCR buffer (Takara). .. The primer sets used for amplification were as follows: Sig-1R forward, 5′-GCCTTCTCTCGTCTGATCGT-3′ and Sig-1R reverse, 5′-GCCAAAGAGGTAGGTGGTGA-3′ β -actin forward, 5′-GTTTGAGACCTCAACACC-3′ and β -actin reverse, 5′-GTGGTGGTGAAGCTTAG-3′, X-box binding protein 1 (XBP-1), 5′-TAAGACAGCGCTTGGGGATGG-3′, and 5′-CAGAATCCATGGGGAGATGTT-3′.

    DNA Extraction:

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: Paragraph title: DNA extraction and mutation analysis of the WASP gene. ... For fluorescent genotyping (GeneScan) analysis, the nucleotide 1150–1358 fragment of the WASP genomic DNA was amplified from 300 ng of DNA extracted from PBMCs, immunomagnetic bead-purified CD3+ T lymphocytes, CD3+ cell–depleted PMNs and sorted CD20+ B lymphocytes, CD56+ NK cells, and CD14+ monocytes, using 20 pmol each of the 5′-TCC AGC TAC TGG ACG TTC TG-3′ forward primer and the FAM-labeled 5′-TTC CCT GCC GGA TTT GAT-3′ reverse primer in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq Buffer, 2 mM Mg2+ , and 200 μM dNTPs (all from Takara Bio Inc., Shiga, Japan).

    Mutagenesis:

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: Paragraph title: DNA extraction and mutation analysis of the WASP gene. ... For fluorescent genotyping (GeneScan) analysis, the nucleotide 1150–1358 fragment of the WASP genomic DNA was amplified from 300 ng of DNA extracted from PBMCs, immunomagnetic bead-purified CD3+ T lymphocytes, CD3+ cell–depleted PMNs and sorted CD20+ B lymphocytes, CD56+ NK cells, and CD14+ monocytes, using 20 pmol each of the 5′-TCC AGC TAC TGG ACG TTC TG-3′ forward primer and the FAM-labeled 5′-TTC CCT GCC GGA TTT GAT-3′ reverse primer in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq Buffer, 2 mM Mg2+ , and 200 μM dNTPs (all from Takara Bio Inc., Shiga, Japan).

    Article Title: A Novel A1088T Mutation in the Glucose-6-Phosphate Dehydrogenase Gene Detected by RT-PCR Combined with DNA Sequencing
    Article Snippet: .. The PCR amplification was performed with EX-Taq polymerase and GC buffer(Takara) in a thermocycler(Biometra, Germany) for an initial denaturing cycle at 94 °C for 2 min, followed by 30 cycles (94 °C for 30 s, 59 °C for 30 s, 72 °C for 2 min), and a final extension step at 72 °C for 8 min. Genomic DNA was used to verify the novel mutation site and analyze mutations in the promoter region (from −509 bp to +26 bp). .. The promoter region of human G6PD DNA was amplified by PCR with primer pairs 5′-CCACGGATGGAACCCTGTC-3′/5′-CCGAAGTGTACGACCGTTTC-3′.

    Article Title: High-Throughput Discovery of Chloroplast and Mitochondrial DNA Polymorphisms in Brassicaceae Species by ORG-EcoTILLING
    Article Snippet: Paragraph title: PCR amplification and mutation detection ... The first PCR amplification was performed in a 15 µL reaction volume using 60 ng DNA template, 1.5 µL 10× Ex-buffer, 0.2 mM dNTPs, 0.75 U Ex-Taq polymerase and 0.25 µL 10 µM each primer [Ex-Taq polymerase and Ex-buffer both purchased from TAKARA Biotechnology (Dalian) CO., LTD.].

    Isolation:

    Article Title: Agrobacterium tumefaciens – Mediated transformation of Woodfordia fruticosa (L.) Kurz
    Article Snippet: 2.7 Molecular confirmation of putatively transformed plants Genomic DNA was isolated from the regenerated hygromycin-resistant (putatively transformed) and control plant leaves by cetyl trimethylammonium bromide (C-TAB) method described by Doyle and Doyle . .. PCR reactions were carried out in 20 μl reaction mixture containing 0.5 units of Ex Taq polymerase and 1× Taq buffer (Takara, Dalian, China), 0.2 mM of each dNTP, 0.5 μM of each primer, and 50 ng of template DNA.

    Article Title: A Novel A1088T Mutation in the Glucose-6-Phosphate Dehydrogenase Gene Detected by RT-PCR Combined with DNA Sequencing
    Article Snippet: Total RNA and genomic DNA were isolated from peripheral blood samples by using QIAamp RNA Blood Mini Kit and FlexiGene DNA Kit (Qiagen, Hilden, Germany) respectively. .. The PCR amplification was performed with EX-Taq polymerase and GC buffer(Takara) in a thermocycler(Biometra, Germany) for an initial denaturing cycle at 94 °C for 2 min, followed by 30 cycles (94 °C for 30 s, 59 °C for 30 s, 72 °C for 2 min), and a final extension step at 72 °C for 8 min. Genomic DNA was used to verify the novel mutation site and analyze mutations in the promoter region (from −509 bp to +26 bp).

    Subcloning:

    Article Title: Display of Bombyx mori Alcohol Dehydrogenases on the Bacillus subtilis Spore Surface to Enhance Enzymatic Activity under Adverse Conditions
    Article Snippet: .. Ex Taq polymerase, restriction enzymes, T4 DNA ligase and the subcloning vector pMD18-T were purchased from TaKaRa (Dalian, China). .. Chemicals are all from Sigma (MO, USA) or a domestic provider in China if not stated otherwise.

    RNA Extraction:

    Article Title: Fluvoxamine alleviates ER stress via induction of Sigma-1 receptor
    Article Snippet: Paragraph title: RNA extraction and semiquantitative RT-PCR ... PCR was performed in 50 μ l containing 0.8 μ M of each primer, 0.2 mM dNTPs, 1.25 U of Ex-Taq polymerase, and 10 × PCR buffer (Takara).

    Titration:

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: For fluorescent genotyping (GeneScan) analysis, the nucleotide 1150–1358 fragment of the WASP genomic DNA was amplified from 300 ng of DNA extracted from PBMCs, immunomagnetic bead-purified CD3+ T lymphocytes, CD3+ cell–depleted PMNs and sorted CD20+ B lymphocytes, CD56+ NK cells, and CD14+ monocytes, using 20 pmol each of the 5′-TCC AGC TAC TGG ACG TTC TG-3′ forward primer and the FAM-labeled 5′-TTC CCT GCC GGA TTT GAT-3′ reverse primer in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq Buffer, 2 mM Mg2+ , and 200 μM dNTPs (all from Takara Bio Inc., Shiga, Japan). .. Titration studies showed that this procedure allowed detection of one copy of the Δ19bp mutation present within 5,000 cells (data not shown).

    Sequencing:

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: Sequencing was performed on gel-purified PCR products or PCR products subcloned in the pCR2.1 vector (Invitrogen Corp., Carlsbad, California, USA) using the ABI Prism BigDye Terminator Cycle sequencing kit on an ABI Prism 310 Genetic Analyzer (Perkin-Elmer Applied Biosystems, Foster City, California, USA), as described previously ( ). .. For fluorescent genotyping (GeneScan) analysis, the nucleotide 1150–1358 fragment of the WASP genomic DNA was amplified from 300 ng of DNA extracted from PBMCs, immunomagnetic bead-purified CD3+ T lymphocytes, CD3+ cell–depleted PMNs and sorted CD20+ B lymphocytes, CD56+ NK cells, and CD14+ monocytes, using 20 pmol each of the 5′-TCC AGC TAC TGG ACG TTC TG-3′ forward primer and the FAM-labeled 5′-TTC CCT GCC GGA TTT GAT-3′ reverse primer in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq Buffer, 2 mM Mg2+ , and 200 μM dNTPs (all from Takara Bio Inc., Shiga, Japan).

    Article Title: A Novel A1088T Mutation in the Glucose-6-Phosphate Dehydrogenase Gene Detected by RT-PCR Combined with DNA Sequencing
    Article Snippet: The PCR amplification was performed with EX-Taq polymerase and GC buffer(Takara) in a thermocycler(Biometra, Germany) for an initial denaturing cycle at 94 °C for 2 min, followed by 30 cycles (94 °C for 30 s, 59 °C for 30 s, 72 °C for 2 min), and a final extension step at 72 °C for 8 min. Genomic DNA was used to verify the novel mutation site and analyze mutations in the promoter region (from −509 bp to +26 bp). .. The PCR amplification was performed with EX-Taq polymerase and GC buffer(Takara) in a thermocycler(Biometra, Germany) for an initial denaturing cycle at 94 °C for 2 min, followed by 30 cycles (94 °C for 30 s, 59 °C for 30 s, 72 °C for 2 min), and a final extension step at 72 °C for 8 min. Genomic DNA was used to verify the novel mutation site and analyze mutations in the promoter region (from −509 bp to +26 bp).

    Article Title: High Prevalence of Plasmid-Mediated Quinolone Resistance Determinants qnr, aac(6?)-Ib-cr, and qepA among Ceftiofur-Resistant Enterobacteriaceae Isolates from Companion and Food-Producing Animals ▿
    Article Snippet: Enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) was performed with all PMQR determinant-positive isolates. .. The PCR amplifications were performed in 25-μl volumes containing 2.5 mM MgCl2 , 0.5 U of Ex Taq polymerase, 0.25 mM each deoxynucleoside triphosphate, 2.5 μl of 10× amplification buffer, 10 ng of crude template DNA, and 25 pmol each of two opposing primers, primers ERIC1R (5′-ATGTAAGCTCCTGGGGATTCA-3′) and ERIC2 (5′-AAGTAAGTGACTGGGGTGAGCG-3′) (TaKaRa Bio, Dalian, China) ( ).

    Purification:

    Article Title: Immune responses of Helicoverpa armigera to different kinds of pathogens
    Article Snippet: .. Chemicals The chemicals were obtained from the following separate companies: Unizol reagent (Biostar Company, Shanghai, China); Reverse Transcriptase (RT) Kit (Promega Biosciences, Madison WI, USA); PCR purification kit (Shengong, Shanghai, China); Ex Taq Polymerase and SYBR® Premix EX Taq (TaKaRa Biotech, Dalian, China); and 4'-6-Diamidino-2-phenylindole dihydrochloride (DAPI, 1 μg/ml in water, San Jose, United States). ..

    Article Title: Molecular Cloning and Characterization of Taurocyamine Kinase from Clonorchis sinensis: A Candidate Chemotherapeutic Target
    Article Snippet: Amplification of 3′-cDNA end of C. sinensis PK D2 domain Polymerase chain reaction (PCR) was carried out with reaction mixture containing cDNA, 10 pmol of each primer, 2 µl of 2.5 mM of dNTPs, 1 U of Ex Taq polymerase, 2.5 µl of 10× Ex Taq buffer (TaKaRa, Tokyo, Japan). .. PCR products were purified using GENE CLEAN Kit (Funakoshi, Tokyo, Japan).

    Plasmid Preparation:

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: Sequencing was performed on gel-purified PCR products or PCR products subcloned in the pCR2.1 vector (Invitrogen Corp., Carlsbad, California, USA) using the ABI Prism BigDye Terminator Cycle sequencing kit on an ABI Prism 310 Genetic Analyzer (Perkin-Elmer Applied Biosystems, Foster City, California, USA), as described previously ( ). .. For fluorescent genotyping (GeneScan) analysis, the nucleotide 1150–1358 fragment of the WASP genomic DNA was amplified from 300 ng of DNA extracted from PBMCs, immunomagnetic bead-purified CD3+ T lymphocytes, CD3+ cell–depleted PMNs and sorted CD20+ B lymphocytes, CD56+ NK cells, and CD14+ monocytes, using 20 pmol each of the 5′-TCC AGC TAC TGG ACG TTC TG-3′ forward primer and the FAM-labeled 5′-TTC CCT GCC GGA TTT GAT-3′ reverse primer in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq Buffer, 2 mM Mg2+ , and 200 μM dNTPs (all from Takara Bio Inc., Shiga, Japan).

    Article Title: Cloning and expression of visfatin and screening of oligopeptides binding with visfatin
    Article Snippet: Prokaryotic expression vector pQE-30Xa and E.coli strains M15 [pREP4] were purchased from Qiagen. .. The enzymes Sal I, Stu I, Pvu II, Ex Taq polymerase and dNTP were purchased from Takara Bio Inc. M-MLV reverse transcriptase and T4 DNA ligase were purchased from Promega.

    Article Title: Display of Bombyx mori Alcohol Dehydrogenases on the Bacillus subtilis Spore Surface to Enhance Enzymatic Activity under Adverse Conditions
    Article Snippet: .. Ex Taq polymerase, restriction enzymes, T4 DNA ligase and the subcloning vector pMD18-T were purchased from TaKaRa (Dalian, China). .. Chemicals are all from Sigma (MO, USA) or a domestic provider in China if not stated otherwise.

    Software:

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: For fluorescent genotyping (GeneScan) analysis, the nucleotide 1150–1358 fragment of the WASP genomic DNA was amplified from 300 ng of DNA extracted from PBMCs, immunomagnetic bead-purified CD3+ T lymphocytes, CD3+ cell–depleted PMNs and sorted CD20+ B lymphocytes, CD56+ NK cells, and CD14+ monocytes, using 20 pmol each of the 5′-TCC AGC TAC TGG ACG TTC TG-3′ forward primer and the FAM-labeled 5′-TTC CCT GCC GGA TTT GAT-3′ reverse primer in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq Buffer, 2 mM Mg2+ , and 200 μM dNTPs (all from Takara Bio Inc., Shiga, Japan). .. One microliter of the amplification product was then combined to 12 μl of deionized formamide and 0.5 μl of GeneScan size standard (Perkin-Elmer Applied Biosystems) and analyzed on a ABI Prism 310 Genetic Analyzer using the GeneScan Analysis Software (Perkin-Elmer Applied Biosystems).

    Article Title: Second-site mutation in the Wiskott-Aldrich syndrome (WAS) protein gene causes somatic mosaicism in two WAS siblings
    Article Snippet: Briefly, 50 ng of DNA extracted from sorted WASP+ and WASP– T lymphocytes was subjected to PCR amplification using a TCR Vβ consensus primer [Vβ pan: 5′-CTC GAA TTC T(T/G) T(A/T) (C/T)T GGT A(C/T) C(G/A)(T/A)CA-3′; 30 pmol] and a TCR-Jβ family-specific primer (Jβ 2.1: 5′-ACG-GTG-AGC-CGT-GTC-CC-3′; 10 pmol) ( ) in a 50-μl volume reaction containing 1.25 U Ex Taq polymerase, 1× Ex Taq buffer, 2 mM Mg2+ , and 200 μM dNTP (all from Takara Bio Inc.). .. One microliter of the second PCR reaction product was combined with 12 μl of deionized formamide and 0.5 μl of GeneScan size standard (Perkin-Elmer Applied Biosystems) and analyzed on an ABI Prism 310 Genetic Analyzer using the GeneScan Analysis Software (Perkin-Elmer Applied Biosystems).

    Negative Control:

    Article Title: High Prevalence of Plasmid-Mediated Quinolone Resistance Determinants qnr, aac(6?)-Ib-cr, and qepA among Ceftiofur-Resistant Enterobacteriaceae Isolates from Companion and Food-Producing Animals ▿
    Article Snippet: The PCR amplifications were performed in 25-μl volumes containing 2.5 mM MgCl2 , 0.5 U of Ex Taq polymerase, 0.25 mM each deoxynucleoside triphosphate, 2.5 μl of 10× amplification buffer, 10 ng of crude template DNA, and 25 pmol each of two opposing primers, primers ERIC1R (5′-ATGTAAGCTCCTGGGGATTCA-3′) and ERIC2 (5′-AAGTAAGTGACTGGGGTGAGCG-3′) (TaKaRa Bio, Dalian, China) ( ). .. A negative control consisting of the same reaction mixture without a DNA template was included in each PCR.

    Positron Emission Tomography:

    Article Title: Display of Bombyx mori Alcohol Dehydrogenases on the Bacillus subtilis Spore Surface to Enhance Enzymatic Activity under Adverse Conditions
    Article Snippet: The expression vector pET-30a(+) and Escherichia coli strains DH5α and BL21(DE3) were obtained from Novagen (CA, USA). .. Ex Taq polymerase, restriction enzymes, T4 DNA ligase and the subcloning vector pMD18-T were purchased from TaKaRa (Dalian, China).

    Agarose Gel Electrophoresis:

    Article Title: Expression of a Functional zipFv Antibody Fragment and Its Fusions with Alkaline Phosphatase in the Cytoplasm of an Escherichia coli
    Article Snippet: DNA cloning procedure All DNA cloning experiments were carried out according to the standard procedures , and Ex-Taq polymerase and Pfu DNA polymerase (Takara, Japan) were successfully used for the polymerase chain reactions (PCR). .. The VLκ gene fragment of SP112 was recovered from 1.2% agarose gel, treated with Sal I and Sac II (Takara), and cloned into pCzFv, pCzFvHAP and pCzFvLAP at a 2 : 1 molar ratio using T4 DNA ligase (Takara) at 16℃ overnight.

    In Vitro:

    Article Title: Agrobacterium tumefaciens – Mediated transformation of Woodfordia fruticosa (L.) Kurz
    Article Snippet: PCR reactions were carried out in 20 μl reaction mixture containing 0.5 units of Ex Taq polymerase and 1× Taq buffer (Takara, Dalian, China), 0.2 mM of each dNTP, 0.5 μM of each primer, and 50 ng of template DNA. .. The PCR conditions for hpt gene detection were set as initial-denaturation at 94 °C for 5 min, 30 cycles of denaturation at 95 °C for 40 s, annealing at 56 °C for 1 min, extension at 72 °C for 40 s and final extension at 72 °C for 15 min. To know the expression of transgenes by RT-PCR (reverse transcription-PCR), total RNA was isolated from in vitro regenerated transgenic plant lines following the established standard protocol and treated with DNase I (Takara, Dalian, China) to remove DNA traces.

    Transgenic Assay:

    Article Title: Agrobacterium tumefaciens – Mediated transformation of Woodfordia fruticosa (L.) Kurz
    Article Snippet: PCR reactions were carried out in 20 μl reaction mixture containing 0.5 units of Ex Taq polymerase and 1× Taq buffer (Takara, Dalian, China), 0.2 mM of each dNTP, 0.5 μM of each primer, and 50 ng of template DNA. .. The PCR conditions for hpt gene detection were set as initial-denaturation at 94 °C for 5 min, 30 cycles of denaturation at 95 °C for 40 s, annealing at 56 °C for 1 min, extension at 72 °C for 40 s and final extension at 72 °C for 15 min. To know the expression of transgenes by RT-PCR (reverse transcription-PCR), total RNA was isolated from in vitro regenerated transgenic plant lines following the established standard protocol and treated with DNase I (Takara, Dalian, China) to remove DNA traces.

    Spectrophotometry:

    Article Title: Fluvoxamine alleviates ER stress via induction of Sigma-1 receptor
    Article Snippet: For each extract, the RNA concentration was determined by spectrophotometry at 260 nm, and 2 μ g of RNA was used with a PrimeScript II 1st Strand cDNA Synthesis Kit (Takara, Shiga, Japan) for the RT reaction. .. PCR was performed in 50 μ l containing 0.8 μ M of each primer, 0.2 mM dNTPs, 1.25 U of Ex-Taq polymerase, and 10 × PCR buffer (Takara).

    Concentration Assay:

    Article Title: Fluvoxamine alleviates ER stress via induction of Sigma-1 receptor
    Article Snippet: For each extract, the RNA concentration was determined by spectrophotometry at 260 nm, and 2 μ g of RNA was used with a PrimeScript II 1st Strand cDNA Synthesis Kit (Takara, Shiga, Japan) for the RT reaction. .. PCR was performed in 50 μ l containing 0.8 μ M of each primer, 0.2 mM dNTPs, 1.25 U of Ex-Taq polymerase, and 10 × PCR buffer (Takara).

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