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TaKaRa ex taq dna polymerase
Ex Taq Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1112 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 1112 article reviews
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ex taq dna polymerase - by Bioz Stars, 2020-04
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Amplification:

Article Title: Molecular analysis of population and De Novo transcriptome sequencing of Thai medaka, Oryzias minutillus (Teleostei: Adrianichthyidae)
Article Snippet: .. The two pairs of primers that were used were 5′–ggRggWATACCWgTTgAACACCC–3′ and 5′–ggTTTRACCCgCAAAAgCCggg–3′ for the D-loop and 5′–CCYCAgggCTgRTAAgAAgAggA–3′ and 5′–ggTACgTgggAgTTWAACCCACAA–3′ for Cox1 with Ex Taq polymerase (Takara, Japan) for DNA target 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: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: .. The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan). .. The PCR parameters were 35 or 28 cycles at 95 °C for 30 s, 60 °C for 1 min and 72 °C for 1 min after hot starting at 95 °C for 3 min, and the products were detected using 1% agarose gel electrophoresis.

Article Title: Newborn Screening for Spinal Muscular Atrophy: DNA Preparation from Dried Blood Spot and DNA Polymerase Selection in PCR
Article Snippet: When TaKaRa Ex Taq™ polymerase was used, DNA extraction process might be essential. .. Without DNA extraction process, PCR amplification was not successful even at 40 cycles.

Article Title: The SEPS1 G-105A Polymorphism Is Associated with Risk of Spontaneous Preterm Birth in a Chinese Population
Article Snippet: PCR was performed with a 25 µL reaction mixture containing approximately 20 ng DNA, 0.2 µM of each primer, 0.2 mM deoxynucleoside triphosphates, and 0.5 unit Ex Taq Polymerase in 1X reaction buffer (Takara BioTech, Dalian, China). .. The amplification reaction was carried out in the following conditions: an initial melting step of 2 min at 95°C, followed by 35 cycles of 30 sec at 94°C, 30 sec at 57°C and 30 sec at 72°C with a final elongation of 7 min at 72°C.

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: Regulatory Role of PlaR (YiaJ) for Plant Utilization in Escherichia coli K-12
Article Snippet: .. DIG-labeled probes were prepared by PCR amplification using BW25113 genomic DNA as template, DIG-11-dUTP (Roche) and dNTP as substrates, gene-specific forward and reverse primers (Table ), and Ex Taq DNA polymerase (Takara). .. Total RNAs (2 μg) were incubated in formaldehyde-MOPS (morpholinepropanesulfonic acid) gel-loading buffer for 10 min at 65 °C for denaturation, subjected to electrophoresis on formaldehyde-containing 2% agarose gel, and then transferred to nylon membrane (Roche).

Article Title: Yalongjiang River Has Had an Important Role in the Dispersal and Divergence of Rosa soulieana in the Hengduan Mountains of China
Article Snippet: Four selective primer pairs: EcoR I (FAM) -AAG/ Mse I -CAT, EcoR I (FAM) -AAG/Mse I- CGT, EcoR I (FAM) -AAG/Mse I- CTC and EcoR I (FAM)-AAG/Mse I–CTT (FAM is a kind of fluorescent dye; primer sequences are listed in ) were selected for the amplification reaction. .. The reaction mix (total volume of 20 μl) included 5 μl 10-fold diluted pre-PCR product, 0.5 U Ex-Taq polymerase, 2 μl 10×PCR buffer (Takara) (Mg2+ free), along with 1.2 μl MgCl2 , 1 μl 10 μM Eco R I Primer (+3), 1 μl 10 μM Mse I Primer (+3), and 1.6 μl 2.5 mM dNTPs (Takara).

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: Identification of zebrafish steroid sulfatase and comparative analysis of the enzymatic properties with human steroid sulfatase
Article Snippet: Takara Ex Taq DNA polymerase was purchased from Clontech Laboratories, Inc. First-strand cDNA synthesis kit was from GE Healthcare Life Sciences. .. Oligonucleotide primers were synthesized by MWG Biotech.

SYBR Green Assay:

Article Title: Molecular Characterization of Viable Legionella spp. in Cooling Tower Water Samples by Combined Use of Ethidium Monoazide and PCR
Article Snippet: .. PCR reaction mixtures (30 μL) contained 5 μL of template DNA, 1 μL of 10 μM forward primer, 1 μL of 10 μM reverse primer, 0.15 μL of Ex Taq polymerase, 2.4 μL of dNTPs, 3 μL of 10×Ex buffer (TaKaRa Bio), and 1 μL of 1,000 dilutions of SYBR Green I dye (Lonza, ME, USA) with dimethyl sulfoxide in a Thermal Cycler Dice Real Time System II (TaKaRa Bio). .. The PCR program parameters were: an initial denaturation step of 2 min at 95°C followed by 45 cycles of denaturation for 15 s at 95°C, annealing for 30 s at 65°C (LEG primer pair) or 50°C (Lmip primer pair), and extension for 60 s at 72°C.

Incubation:

Article Title: Regulatory Role of PlaR (YiaJ) for Plant Utilization in Escherichia coli K-12
Article Snippet: DIG-labeled probes were prepared by PCR amplification using BW25113 genomic DNA as template, DIG-11-dUTP (Roche) and dNTP as substrates, gene-specific forward and reverse primers (Table ), and Ex Taq DNA polymerase (Takara). .. Total RNAs (2 μg) were incubated in formaldehyde-MOPS (morpholinepropanesulfonic acid) gel-loading buffer for 10 min at 65 °C for denaturation, subjected to electrophoresis on formaldehyde-containing 2% agarose gel, and then transferred to nylon membrane (Roche).

Article Title: Yalongjiang River Has Had an Important Role in the Dispersal and Divergence of Rosa soulieana in the Hengduan Mountains of China
Article Snippet: After an initial incubation at 65°C for 5 min, 30 cycles of 94°C for 30 s, 56°C for 30 s and 72°C for 1 min were performed with a final extension for 5 min. We electrophoresed 5 μl pre-amplified product on a 1.0% agarose gel to identify the most variable selective primer extensions. .. The reaction mix (total volume of 20 μl) included 5 μl 10-fold diluted pre-PCR product, 0.5 U Ex-Taq polymerase, 2 μl 10×PCR buffer (Takara) (Mg2+ free), along with 1.2 μl MgCl2 , 1 μl 10 μM Eco R I Primer (+3), 1 μl 10 μM Mse I Primer (+3), and 1.6 μl 2.5 mM dNTPs (Takara).

Expressing:

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: Identification of zebrafish steroid sulfatase and comparative analysis of the enzymatic properties with human steroid sulfatase
Article Snippet: Trizol and pcDNA4 mammalian expression vector were products of Invitrogen. .. Takara Ex Taq DNA polymerase was purchased from Clontech Laboratories, Inc. First-strand cDNA synthesis kit was from GE Healthcare Life Sciences.

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.

Transformation Assay:

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: Regulatory Role of PlaR (YiaJ) for Plant Utilization in Escherichia coli K-12
Article Snippet: DIG-labeled probes were prepared by PCR amplification using BW25113 genomic DNA as template, DIG-11-dUTP (Roche) and dNTP as substrates, gene-specific forward and reverse primers (Table ), and Ex Taq DNA polymerase (Takara). .. Hybridization was performed with DIG easy Hyb system (Roche) at 50 °C overnight with a DIG-labeled probe.

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).

Northern Blot:

Article Title: Regulatory Role of PlaR (YiaJ) for Plant Utilization in Escherichia coli K-12
Article Snippet: Paragraph title: Northern blotting assay ... DIG-labeled probes were prepared by PCR amplification using BW25113 genomic DNA as template, DIG-11-dUTP (Roche) and dNTP as substrates, gene-specific forward and reverse primers (Table ), and Ex Taq DNA polymerase (Takara).

Generated:

Article Title: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: The first-strand cDNA was generated from 4 µg of total RNA of each using Ready-To-Go You-Prime First-Strand Beads (GE Healthcare, Buckinghamshire, UK) with an oligo dT primer. .. The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan).

DNA Sequencing:

Article Title: Molecular analysis of population and De Novo transcriptome sequencing of Thai medaka, Oryzias minutillus (Teleostei: Adrianichthyidae)
Article Snippet: The two pairs of primers that were used were 5′–ggRggWATACCWgTTgAACACCC–3′ and 5′–ggTTTRACCCgCAAAAgCCggg–3′ for the D-loop and 5′–CCYCAgggCTgRTAAgAAgAggA–3′ and 5′–ggTACgTgggAgTTWAACCCACAA–3′ for Cox1 with Ex Taq polymerase (Takara, Japan) for DNA target amplification. .. DNA sequencing was performed using an automated DNA analyzer ABI 3730xl system (Applied Biosystems, US).

Reverse Transcription Polymerase Chain Reaction:

Article Title: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: Paragraph title: RT-PCR ... The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan).

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: 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.

Recombinant:

Article Title: Identification of zebrafish steroid sulfatase and comparative analysis of the enzymatic properties with human steroid sulfatase
Article Snippet: Recombinant SULT1E1, SULT2A1, and SULT2Blb were prepared using the pGEX-2TK expression and purification system as previously described [ , ]. .. Takara Ex Taq DNA polymerase was purchased from Clontech Laboratories, Inc. First-strand cDNA synthesis kit was from GE Healthcare Life Sciences.

DNA Extraction:

Article Title: Molecular analysis of population and De Novo transcriptome sequencing of Thai medaka, Oryzias minutillus (Teleostei: Adrianichthyidae)
Article Snippet: The caudal fins were cut and stored in absolute ethanol at –20 °C prior to DNA extraction. .. The two pairs of primers that were used were 5′–ggRggWATACCWgTTgAACACCC–3′ and 5′–ggTTTRACCCgCAAAAgCCggg–3′ for the D-loop and 5′–CCYCAgggCTgRTAAgAAgAggA–3′ and 5′–ggTACgTgggAgTTWAACCCACAA–3′ for Cox1 with Ex Taq polymerase (Takara, Japan) for DNA target amplification.

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: Newborn Screening for Spinal Muscular Atrophy: DNA Preparation from Dried Blood Spot and DNA Polymerase Selection in PCR
Article Snippet: .. When TaKaRa Ex Taq™ polymerase was used, DNA extraction process might be essential. .. Without DNA extraction process, PCR amplification was not successful even at 40 cycles.

Article Title: Yalongjiang River Has Had an Important Role in the Dispersal and Divergence of Rosa soulieana in the Hengduan Mountains of China
Article Snippet: Paragraph title: DNA extraction and AFLP genotyping ... The reaction mix (total volume of 20 μl) included 5 μl 10-fold diluted pre-PCR product, 0.5 U Ex-Taq polymerase, 2 μl 10×PCR buffer (Takara) (Mg2+ free), along with 1.2 μl MgCl2 , 1 μl 10 μM Eco R I Primer (+3), 1 μl 10 μM Mse I Primer (+3), and 1.6 μl 2.5 mM dNTPs (Takara).

Nucleic Acid Electrophoresis:

Article Title: Newborn Screening for Spinal Muscular Atrophy: DNA Preparation from Dried Blood Spot and DNA Polymerase Selection in PCR
Article Snippet: When TaKaRa Ex Taq™ polymerase was used, DNA extraction process might be essential. .. Whereas, when KOD FX Neo™ was used, PCR amplification was successful without DNA extraction process; PCR products at both 35 and 40 cycles were clearly detected on the gel electrophoresis ( ).

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).

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.

Size-exclusion Chromatography:

Article Title: The SEPS1 G-105A Polymorphism Is Associated with Risk of Spontaneous Preterm Birth in a Chinese Population
Article Snippet: PCR was performed with a 25 µL reaction mixture containing approximately 20 ng DNA, 0.2 µM of each primer, 0.2 mM deoxynucleoside triphosphates, and 0.5 unit Ex Taq Polymerase in 1X reaction buffer (Takara BioTech, Dalian, China). .. The amplification reaction was carried out in the following conditions: an initial melting step of 2 min at 95°C, followed by 35 cycles of 30 sec at 94°C, 30 sec at 57°C and 30 sec at 72°C with a final elongation of 7 min at 72°C.

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).

Polymerase Chain Reaction:

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: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan). .. The PCR parameters were 35 or 28 cycles at 95 °C for 30 s, 60 °C for 1 min and 72 °C for 1 min after hot starting at 95 °C for 3 min, and the products were detected using 1% agarose gel electrophoresis.

Article Title: Molecular Characterization of Viable Legionella spp. in Cooling Tower Water Samples by Combined Use of Ethidium Monoazide and PCR
Article Snippet: .. PCR reaction mixtures (30 μL) contained 5 μL of template DNA, 1 μL of 10 μM forward primer, 1 μL of 10 μM reverse primer, 0.15 μL of Ex Taq polymerase, 2.4 μL of dNTPs, 3 μL of 10×Ex buffer (TaKaRa Bio), and 1 μL of 1,000 dilutions of SYBR Green I dye (Lonza, ME, USA) with dimethyl sulfoxide in a Thermal Cycler Dice Real Time System II (TaKaRa Bio). .. The PCR program parameters were: an initial denaturation step of 2 min at 95°C followed by 45 cycles of denaturation for 15 s at 95°C, annealing for 30 s at 65°C (LEG primer pair) or 50°C (Lmip primer pair), and extension for 60 s at 72°C.

Article Title: Newborn Screening for Spinal Muscular Atrophy: DNA Preparation from Dried Blood Spot and DNA Polymerase Selection in PCR
Article Snippet: Here, we investigated whether PCR can be performed with or without DNA extraction process from DBS. .. When TaKaRa Ex Taq™ polymerase was used, DNA extraction process might be essential.

Article Title: The SEPS1 G-105A Polymorphism Is Associated with Risk of Spontaneous Preterm Birth in a Chinese Population
Article Snippet: .. PCR was performed with a 25 µL reaction mixture containing approximately 20 ng DNA, 0.2 µM of each primer, 0.2 mM deoxynucleoside triphosphates, and 0.5 unit Ex Taq Polymerase in 1X reaction buffer (Takara BioTech, Dalian, China). .. The amplification reaction was carried out in the following conditions: an initial melting step of 2 min at 95°C, followed by 35 cycles of 30 sec at 94°C, 30 sec at 57°C and 30 sec at 72°C with a final elongation of 7 min at 72°C.

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: 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: Regulatory Role of PlaR (YiaJ) for Plant Utilization in Escherichia coli K-12
Article Snippet: .. DIG-labeled probes were prepared by PCR amplification using BW25113 genomic DNA as template, DIG-11-dUTP (Roche) and dNTP as substrates, gene-specific forward and reverse primers (Table ), and Ex Taq DNA polymerase (Takara). .. Total RNAs (2 μg) were incubated in formaldehyde-MOPS (morpholinepropanesulfonic acid) gel-loading buffer for 10 min at 65 °C for denaturation, subjected to electrophoresis on formaldehyde-containing 2% agarose gel, and then transferred to nylon membrane (Roche).

Article Title: Yalongjiang River Has Had an Important Role in the Dispersal and Divergence of Rosa soulieana in the Hengduan Mountains of China
Article Snippet: A pre-selective polymerase chain reaction (PCR) was done using a Bio-Rad machine with a single selective nucleotide extension. .. The reaction mix (total volume of 20 μl) included 5 μl 10-fold diluted pre-PCR product, 0.5 U Ex-Taq polymerase, 2 μl 10×PCR buffer (Takara) (Mg2+ free), along with 1.2 μl MgCl2 , 1 μl 10 μM Eco R I Primer (+3), 1 μl 10 μM Mse I Primer (+3), and 1.6 μl 2.5 mM dNTPs (Takara).

Gel Extraction:

Article Title: Molecular analysis of population and De Novo transcriptome sequencing of Thai medaka, Oryzias minutillus (Teleostei: Adrianichthyidae)
Article Snippet: The two pairs of primers that were used were 5′–ggRggWATACCWgTTgAACACCC–3′ and 5′–ggTTTRACCCgCAAAAgCCggg–3′ for the D-loop and 5′–CCYCAgggCTgRTAAgAAgAggA–3′ and 5′–ggTACgTgggAgTTWAACCCACAA–3′ for Cox1 with Ex Taq polymerase (Takara, Japan) for DNA target amplification. .. The PCR products were viewed under the UV transillumination of 1% agarose gels, were stained with SYBR® Safe DNA Gel Stain (Thermo Fisher Scientific, US) and were extracted from those gels by using the QIAquick Gel Extraction Kit (Qiagen, Germany).

Chloramphenicol Acetyltransferase Assay:

Article Title: Yalongjiang River Has Had an Important Role in the Dispersal and Divergence of Rosa soulieana in the Hengduan Mountains of China
Article Snippet: Four selective primer pairs: EcoR I (FAM) -AAG/ Mse I -CAT, EcoR I (FAM) -AAG/Mse I- CGT, EcoR I (FAM) -AAG/Mse I- CTC and EcoR I (FAM)-AAG/Mse I–CTT (FAM is a kind of fluorescent dye; primer sequences are listed in ) were selected for the amplification reaction. .. The reaction mix (total volume of 20 μl) included 5 μl 10-fold diluted pre-PCR product, 0.5 U Ex-Taq polymerase, 2 μl 10×PCR buffer (Takara) (Mg2+ free), along with 1.2 μl MgCl2 , 1 μl 10 μM Eco R I Primer (+3), 1 μl 10 μM Mse I Primer (+3), and 1.6 μl 2.5 mM dNTPs (Takara).

Purification:

Article Title: Identification of zebrafish steroid sulfatase and comparative analysis of the enzymatic properties with human steroid sulfatase
Article Snippet: Recombinant SULT1E1, SULT2A1, and SULT2Blb were prepared using the pGEX-2TK expression and purification system as previously described [ , ]. .. Takara Ex Taq DNA polymerase was purchased from Clontech Laboratories, Inc. First-strand cDNA synthesis kit was from GE Healthcare Life Sciences.

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: Identification of zebrafish steroid sulfatase and comparative analysis of the enzymatic properties with human steroid sulfatase
Article Snippet: Trizol and pcDNA4 mammalian expression vector were products of Invitrogen. .. Takara Ex Taq DNA polymerase was purchased from Clontech Laboratories, Inc. First-strand cDNA synthesis kit was from GE Healthcare Life Sciences.

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).

Real-time Polymerase Chain Reaction:

Article Title: Molecular Characterization of Viable Legionella spp. in Cooling Tower Water Samples by Combined Use of Ethidium Monoazide and PCR
Article Snippet: Populations of Legionella spp. and L. pneumophila were quantified by qPCR using the primer pairs LEG-225F (5′-AAG ATT AGC CTG CGT CCG AT-3′) and LEG-858R (5′-GTC AAC TTA TCG CGT TTG CT-3′) , and Lmip L920 (5′-GCT ACA GAC AAG GAT AAG TTG-3′) and Lmip R1548 (5′-GTT TTG TAT GAC TTT AAT TCA-3′) , respectively. .. PCR reaction mixtures (30 μL) contained 5 μL of template DNA, 1 μL of 10 μM forward primer, 1 μL of 10 μM reverse primer, 0.15 μL of Ex Taq polymerase, 2.4 μL of dNTPs, 3 μL of 10×Ex buffer (TaKaRa Bio), and 1 μL of 1,000 dilutions of SYBR Green I dye (Lonza, ME, USA) with dimethyl sulfoxide in a Thermal Cycler Dice Real Time System II (TaKaRa Bio).

Selection:

Article Title: Molecular Characterization of Viable Legionella spp. in Cooling Tower Water Samples by Combined Use of Ethidium Monoazide and PCR
Article Snippet: A Viable Legionella Selection Kit for PCR ver.2.0 (TaKaRa Bio) including the EMA treatment was used for the clone library construction, as described by the manufacturer. .. PCR reaction mixtures (30 μL) contained 5 μL of template DNA, 1 μL of 10 μM forward primer, 1 μL of 10 μM reverse primer, 0.15 μL of Ex Taq polymerase, 2.4 μL of dNTPs, 3 μL of 10×Ex buffer (TaKaRa Bio), and 1 μL of 1,000 dilutions of SYBR Green I dye (Lonza, ME, USA) with dimethyl sulfoxide in a Thermal Cycler Dice Real Time System II (TaKaRa Bio).

Agarose Gel Electrophoresis:

Article Title: Deficiency of a pyrroline-5-carboxylate reductase produces the yellowish green cocoon ‘Ryokuken’ of the silkworm, Bombyx mori
Article Snippet: The region from transcription initiation to terminator (1F-9063R) and to the 1st intronic region on the Daizo genome (1F-5924R) of BmP5CR1 was amplified by 2.5 U Ex Taq polymerase or LA Taq polymerase (TaKaRa, Japan). .. The PCR parameters were 35 or 28 cycles at 95 °C for 30 s, 60 °C for 1 min and 72 °C for 1 min after hot starting at 95 °C for 3 min, and the products were detected using 1% agarose gel electrophoresis.

Article Title: The SEPS1 G-105A Polymorphism Is Associated with Risk of Spontaneous Preterm Birth in a Chinese Population
Article Snippet: PCR was performed with a 25 µL reaction mixture containing approximately 20 ng DNA, 0.2 µM of each primer, 0.2 mM deoxynucleoside triphosphates, and 0.5 unit Ex Taq Polymerase in 1X reaction buffer (Takara BioTech, Dalian, China). .. An aliquot (5 µL) of PCR product was digested with 4 unit of 4 Msc I (New England Biolabs) and separated on a 2.5% agarose gel.

Article Title: Regulatory Role of PlaR (YiaJ) for Plant Utilization in Escherichia coli K-12
Article Snippet: RNA purity was checked by electrophoresis on 1.5% agarose gel in the presence of formaldehyde followed by staining with Methylene blue. .. DIG-labeled probes were prepared by PCR amplification using BW25113 genomic DNA as template, DIG-11-dUTP (Roche) and dNTP as substrates, gene-specific forward and reverse primers (Table ), and Ex Taq DNA polymerase (Takara).

Article Title: Yalongjiang River Has Had an Important Role in the Dispersal and Divergence of Rosa soulieana in the Hengduan Mountains of China
Article Snippet: After an initial incubation at 65°C for 5 min, 30 cycles of 94°C for 30 s, 56°C for 30 s and 72°C for 1 min were performed with a final extension for 5 min. We electrophoresed 5 μl pre-amplified product on a 1.0% agarose gel to identify the most variable selective primer extensions. .. The reaction mix (total volume of 20 μl) included 5 μl 10-fold diluted pre-PCR product, 0.5 U Ex-Taq polymerase, 2 μl 10×PCR buffer (Takara) (Mg2+ free), along with 1.2 μl MgCl2 , 1 μl 10 μM Eco R I Primer (+3), 1 μl 10 μM Mse I Primer (+3), and 1.6 μl 2.5 mM dNTPs (Takara).

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.

Electrophoresis:

Article Title: Regulatory Role of PlaR (YiaJ) for Plant Utilization in Escherichia coli K-12
Article Snippet: RNA purity was checked by electrophoresis on 1.5% agarose gel in the presence of formaldehyde followed by staining with Methylene blue. .. DIG-labeled probes were prepared by PCR amplification using BW25113 genomic DNA as template, DIG-11-dUTP (Roche) and dNTP as substrates, gene-specific forward and reverse primers (Table ), and Ex Taq DNA polymerase (Takara).

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.

CTG Assay:

Article Title: Molecular Characterization of Viable Legionella spp. in Cooling Tower Water Samples by Combined Use of Ethidium Monoazide and PCR
Article Snippet: Populations of Legionella spp. and L. pneumophila were quantified by qPCR using the primer pairs LEG-225F (5′-AAG ATT AGC CTG CGT CCG AT-3′) and LEG-858R (5′-GTC AAC TTA TCG CGT TTG CT-3′) , and Lmip L920 (5′-GCT ACA GAC AAG GAT AAG TTG-3′) and Lmip R1548 (5′-GTT TTG TAT GAC TTT AAT TCA-3′) , respectively. .. PCR reaction mixtures (30 μL) contained 5 μL of template DNA, 1 μL of 10 μM forward primer, 1 μL of 10 μM reverse primer, 0.15 μL of Ex Taq polymerase, 2.4 μL of dNTPs, 3 μL of 10×Ex buffer (TaKaRa Bio), and 1 μL of 1,000 dilutions of SYBR Green I dye (Lonza, ME, USA) with dimethyl sulfoxide in a Thermal Cycler Dice Real Time System II (TaKaRa Bio).

Staining:

Article Title: Molecular analysis of population and De Novo transcriptome sequencing of Thai medaka, Oryzias minutillus (Teleostei: Adrianichthyidae)
Article Snippet: The two pairs of primers that were used were 5′–ggRggWATACCWgTTgAACACCC–3′ and 5′–ggTTTRACCCgCAAAAgCCggg–3′ for the D-loop and 5′–CCYCAgggCTgRTAAgAAgAggA–3′ and 5′–ggTACgTgggAgTTWAACCCACAA–3′ for Cox1 with Ex Taq polymerase (Takara, Japan) for DNA target amplification. .. The PCR products were viewed under the UV transillumination of 1% agarose gels, were stained with SYBR® Safe DNA Gel Stain (Thermo Fisher Scientific, US) and were extracted from those gels by using the QIAquick Gel Extraction Kit (Qiagen, Germany).

Article Title: Regulatory Role of PlaR (YiaJ) for Plant Utilization in Escherichia coli K-12
Article Snippet: RNA purity was checked by electrophoresis on 1.5% agarose gel in the presence of formaldehyde followed by staining with Methylene blue. .. DIG-labeled probes were prepared by PCR amplification using BW25113 genomic DNA as template, DIG-11-dUTP (Roche) and dNTP as substrates, gene-specific forward and reverse primers (Table ), and Ex Taq DNA polymerase (Takara).

Fluorescence In Situ Hybridization:

Article Title: Molecular analysis of population and De Novo transcriptome sequencing of Thai medaka, Oryzias minutillus (Teleostei: Adrianichthyidae)
Article Snippet: The genomic DNA of the fish fin was extracted by using the DNeasy Blood & Tissue Kit (Qiagen, Germany) according to the manufacturer's instructions. .. The two pairs of primers that were used were 5′–ggRggWATACCWgTTgAACACCC–3′ and 5′–ggTTTRACCCgCAAAAgCCggg–3′ for the D-loop and 5′–CCYCAgggCTgRTAAgAAgAggA–3′ and 5′–ggTACgTgggAgTTWAACCCACAA–3′ for Cox1 with Ex Taq polymerase (Takara, Japan) for DNA target amplification.

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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
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    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 330 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa sybr premix ex taq ii
    Quantitative analysis of NOX2 and NOX4 mRNA expression in human OS cell lines. Quantitative analysis of the mRNA expression of NOX family proteins in five human OS cell lines was performed by RT-qPCR using a <t>SYBR</t> Green reagent. The Cq value during the exponential phase of amplification was determined by the real-time monitoring of the fluorescent emission of <t>Taq</t> polymerase nuclease activity. β- actin was used as an internal control for normalization. qPCR efficiencies of the target ( NOX2 and NOX4 ) and control (β-actin) proteins were approximately equal over a concentration range of 0.1–200 ng total cDNA. The relative transcripts were determined by the formula: 1/2 (Cq target-Cq control) ). The bars represent the mean (n=2). OS, osteosarcoma; NOX, NADPH oxidase; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; Cq, quantification cycle; PBMCs, peripheral blood mononuclear cells.
    Sybr Premix Ex Taq Ii, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1391 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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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

    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

    cDNA sequence of mRNA de novo transcribed from TNFSF9 in HepG2 clone 9-1 (A) HepG2 clone 9-1 cells were incubated with 500 ng/ml LPS for the indicated times. cDNA was synthesized from total RNA, and TNFSF9 expression was quantified by real-time PCR as described in “Materials and Methods”. (B) HepG2 cells were incubated with 500 ng/ml LPS in the presence of 5-ethynyl uridine (EU) for 16 hrs. EU-labeled RNA was isolated as described in “Materials and Methods”. TNFSF9 cDNA was amplified by PCR and the PCR products were isolated and sequenced. This experiment is representative of four independent experiments.

    Journal: Molecules and Cells

    Article Title: Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

    doi: 10.14348/molcells.2018.0209

    Figure Lengend Snippet: cDNA sequence of mRNA de novo transcribed from TNFSF9 in HepG2 clone 9-1 (A) HepG2 clone 9-1 cells were incubated with 500 ng/ml LPS for the indicated times. cDNA was synthesized from total RNA, and TNFSF9 expression was quantified by real-time PCR as described in “Materials and Methods”. (B) HepG2 cells were incubated with 500 ng/ml LPS in the presence of 5-ethynyl uridine (EU) for 16 hrs. EU-labeled RNA was isolated as described in “Materials and Methods”. TNFSF9 cDNA was amplified by PCR and the PCR products were isolated and sequenced. This experiment is representative of four independent experiments.

    Article Snippet: Real-Time PCR amplification of TNFSF9 cDNA Real-time PCR analysis (MiniOpticon, Bio Rad, USA) was performed using PCR Master Mix (Takara SYBR® PreMix Ex Taq II) to quantify expression of TNFSF9 mRNA (normalized to β-actin expression).

    Techniques: Sequencing, Incubation, Synthesized, Expressing, Real-time Polymerase Chain Reaction, Labeling, Isolation, Amplification, Polymerase Chain Reaction

    DNA sequence of genomic TNFSF9 gene edited by CRISPR-Cas9 The TNFSF9 genes in genomic DNAs from wild type and mutated HepG2 cells were amplified by PCR. The PCR products were isolated and sequenced as described in “Materials and Methods”. Left panels are chromatograms of the genomic TNFSF9 sequence. On the right are shown the DNA sequences around the region of the mutated triplet in the wildtype and mutant clones. This is one representative of five independent experiments.

    Journal: Molecules and Cells

    Article Title: Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

    doi: 10.14348/molcells.2018.0209

    Figure Lengend Snippet: DNA sequence of genomic TNFSF9 gene edited by CRISPR-Cas9 The TNFSF9 genes in genomic DNAs from wild type and mutated HepG2 cells were amplified by PCR. The PCR products were isolated and sequenced as described in “Materials and Methods”. Left panels are chromatograms of the genomic TNFSF9 sequence. On the right are shown the DNA sequences around the region of the mutated triplet in the wildtype and mutant clones. This is one representative of five independent experiments.

    Article Snippet: Real-Time PCR amplification of TNFSF9 cDNA Real-time PCR analysis (MiniOpticon, Bio Rad, USA) was performed using PCR Master Mix (Takara SYBR® PreMix Ex Taq II) to quantify expression of TNFSF9 mRNA (normalized to β-actin expression).

    Techniques: Sequencing, CRISPR, Amplification, Polymerase Chain Reaction, Isolation, Mutagenesis, Clone Assay

    Heteroduplex formation by the cDNA of de novo transcripts of TNFSF9 from HepG2 clone 9-1 Total RNA and EU-labeled RNAs were isolated from clone 9-1 HepG2 cells. cDNA was synthesized and the TNFSF9 cDNA was amplified by PCR as described in “Materials and Methods”. The PCR products were annealed and incubated in the presence or absence of T7E1 endonuclease, and the products were fractionated on 1.2% agarose gels and visualized under UV. This is one representative of three independent experiments.

    Journal: Molecules and Cells

    Article Title: Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

    doi: 10.14348/molcells.2018.0209

    Figure Lengend Snippet: Heteroduplex formation by the cDNA of de novo transcripts of TNFSF9 from HepG2 clone 9-1 Total RNA and EU-labeled RNAs were isolated from clone 9-1 HepG2 cells. cDNA was synthesized and the TNFSF9 cDNA was amplified by PCR as described in “Materials and Methods”. The PCR products were annealed and incubated in the presence or absence of T7E1 endonuclease, and the products were fractionated on 1.2% agarose gels and visualized under UV. This is one representative of three independent experiments.

    Article Snippet: Real-Time PCR amplification of TNFSF9 cDNA Real-time PCR analysis (MiniOpticon, Bio Rad, USA) was performed using PCR Master Mix (Takara SYBR® PreMix Ex Taq II) to quantify expression of TNFSF9 mRNA (normalized to β-actin expression).

    Techniques: Labeling, Isolation, Synthesized, Amplification, Polymerase Chain Reaction, Incubation

    cDNA sequence of mRNA transcribed from mutant genomic TNFSF9 TNFSF9 cDNA was synthesized and amplified by PCR from total RNA extracted from wild type and mutated HepG2 cells. The PCR products were isolated and sequenced as described in “Materials and Methods”. Left panels are chromatograms of the TNFSF9 cDNA sequences. On the right are shown the cDNA sequences around the region of the mutated triplet in the wild type and mutant clones. This is one representative of four independent experiments.

    Journal: Molecules and Cells

    Article Title: Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

    doi: 10.14348/molcells.2018.0209

    Figure Lengend Snippet: cDNA sequence of mRNA transcribed from mutant genomic TNFSF9 TNFSF9 cDNA was synthesized and amplified by PCR from total RNA extracted from wild type and mutated HepG2 cells. The PCR products were isolated and sequenced as described in “Materials and Methods”. Left panels are chromatograms of the TNFSF9 cDNA sequences. On the right are shown the cDNA sequences around the region of the mutated triplet in the wild type and mutant clones. This is one representative of four independent experiments.

    Article Snippet: Real-Time PCR amplification of TNFSF9 cDNA Real-time PCR analysis (MiniOpticon, Bio Rad, USA) was performed using PCR Master Mix (Takara SYBR® PreMix Ex Taq II) to quantify expression of TNFSF9 mRNA (normalized to β-actin expression).

    Techniques: Sequencing, Mutagenesis, Synthesized, Amplification, Polymerase Chain Reaction, Isolation, Clone Assay

    Quantitative analysis of NOX2 and NOX4 mRNA expression in human OS cell lines. Quantitative analysis of the mRNA expression of NOX family proteins in five human OS cell lines was performed by RT-qPCR using a SYBR Green reagent. The Cq value during the exponential phase of amplification was determined by the real-time monitoring of the fluorescent emission of Taq polymerase nuclease activity. β- actin was used as an internal control for normalization. qPCR efficiencies of the target ( NOX2 and NOX4 ) and control (β-actin) proteins were approximately equal over a concentration range of 0.1–200 ng total cDNA. The relative transcripts were determined by the formula: 1/2 (Cq target-Cq control) ). The bars represent the mean (n=2). OS, osteosarcoma; NOX, NADPH oxidase; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; Cq, quantification cycle; PBMCs, peripheral blood mononuclear cells.

    Journal: Oncology Letters

    Article Title: Inhibition of NADPH oxidase 2 induces apoptosis in osteosarcoma: The role of reactive oxygen species in cell proliferation

    doi: 10.3892/ol.2018.8291

    Figure Lengend Snippet: Quantitative analysis of NOX2 and NOX4 mRNA expression in human OS cell lines. Quantitative analysis of the mRNA expression of NOX family proteins in five human OS cell lines was performed by RT-qPCR using a SYBR Green reagent. The Cq value during the exponential phase of amplification was determined by the real-time monitoring of the fluorescent emission of Taq polymerase nuclease activity. β- actin was used as an internal control for normalization. qPCR efficiencies of the target ( NOX2 and NOX4 ) and control (β-actin) proteins were approximately equal over a concentration range of 0.1–200 ng total cDNA. The relative transcripts were determined by the formula: 1/2 (Cq target-Cq control) ). The bars represent the mean (n=2). OS, osteosarcoma; NOX, NADPH oxidase; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; Cq, quantification cycle; PBMCs, peripheral blood mononuclear cells.

    Article Snippet: RT-qPCR was performed with SYBR Premix Ex Taq II (Takara Bio, Inc., Otsu, Shiga, Japan) in an ABI PRISM 7500 Sequence Detection system (Applied Biosystems; Thermo Fisher Scientific, Inc.).

    Techniques: Expressing, Quantitative RT-PCR, SYBR Green Assay, Amplification, Activity Assay, Real-time Polymerase Chain Reaction, Concentration Assay