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TaKaRa la taq buffer
La Taq Buffer, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 20 article reviews
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la taq buffer - by Bioz Stars, 2020-01
93/100 stars

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Paired-end Tag:

Article Title: Identification of medium-sized genomic deletions with low coverage, mate-paired restricted tags
Article Snippet: Validation by PCR amplification and clone sequencing PCR primers were designed based on the ditags using Primer Premier 5 software (Additional file : Table S3). .. PCR reactions included ~5 ng of genomic DNA, 2 μL of forward primer (10 μM), 2 μL of reverse primer (10 μM), 5 μL of 10× LA Taq Buffer (Takara ), 8 μL of dNTP (2.5 mM), 2 μL of LA Taq polymerase (Takara ) and nuclease-free water to reach a volume of 50 μL.

Amplification:

Article Title: De Novo Assembly of Coding Sequences of the Mangrove Palm (Nypa fruticans) Using RNA-Seq and Discovery of Whole-Genome Duplications in the Ancestor of Palms
Article Snippet: Each reaction mixture contained 0.4 μM each forward and reverse primer, 0.4 mM each dNTP, 1.5 U LA Taq DNA polymerase and 3μl 10x LA Taq buffer (TaKaRa, Japan) and 50–100 ng of genomic DNA as a template. .. PCR amplification was carried out using a Dyad Disciple thermal Cycler (Bio-Rad, Beijing, USA) with the following cycling conditions: pre-denaturation at 94°C for 5 min followed by 33 cycles of 94°C for 30 sec, 57°C for 45 sec and 72°C for 1 min 30 sec, and finally, 10 min at 72°C for extension.

Article Title: Identification of medium-sized genomic deletions with low coverage, mate-paired restricted tags
Article Snippet: Paragraph title: Validation by PCR amplification and clone sequencing ... PCR reactions included ~5 ng of genomic DNA, 2 μL of forward primer (10 μM), 2 μL of reverse primer (10 μM), 5 μL of 10× LA Taq Buffer (Takara ), 8 μL of dNTP (2.5 mM), 2 μL of LA Taq polymerase (Takara ) and nuclease-free water to reach a volume of 50 μL.

Article Title: Ancient Occasional Host Switching of Maternally Transmitted Bacterial Symbionts of Chemosynthetic Vesicomyid Clams
Article Snippet: .. The mitochondrial or symbiont gene fragments were amplified in 25-µL PCR reactions that contained ∼100 ng DNA template, 1× LA-Taq buffer (Mg2+ plus; TaKaRa Bio, Inc., Shiga, Japan), 0.25 mM each deoxynucleotide (dNTP) solution, 0.2 µM each primer, and 0.625 U LA-Taq polymerase (TaKaRa Bio, Inc.). .. The PCR reaction mixtures were incubated under the conditions described in table 2 after the initial denaturation step (96 °C for 2 min), using a TaKara TP600 Thermal cycler (TaKaRa Bio, Inc.).

Article Title: Integrating nested PCR with high-throughput sequencing to characterize mutations of HBV genome in low viral load samples
Article Snippet: Paragraph title: Primers and amplification ... The reaction contained 2 μL of template DNA, 0.5 μL of each 10 μM primer, 2.5 μL of 10× LA Taq Buffer (MgCl2 -free), 4 μL of dNTP mixture (2.5 mM each), 2.5 μL of 25 mM MgCl2 , and 0.25 μL of TaKaRa LA Taq (5 units/μL).

Article Title: Loss of genes related to Nucleotide Excision Repair (NER) and implications for reductive genome evolution in symbionts of deep-sea vesicomyid clams
Article Snippet: Paragraph title: Amplification and sequencing of NER genes ... For the PCR, 25 μl of the reaction mixture contained 80 ng of template DNA, 2.5 μl of 10× LA Taq buffer (TaKaRa), 2.5 μl of dNTP mix (TaKaRa), 5 μl each of the 1 pmol/μl forward and reverse primer solutions, 0.13 μl of LA Taq polymerase solution (TaKaRa) and 8.87 μl of pure water was used.

Article Title: Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera
Article Snippet: The last cycle was followed by final extension at 72°C for 10 min. Each 50 μL PCR reaction contained 2 μL of cDNA template, 5 μL of 10× LA Taq buffer (Mg2+ Free), 4 μL of MgCl2 (25 mM), 4 μL of dNTP mixture (2.5 mM/each), 1 μL of forward and 1 μL of reverse primers (10 μM), 0.5 μL of LA Taq polymerase (Takara Bio.) (5 U/μL) and 32.5 μL of double distilled H2 O. .. The 5’- and 3’-RACE Ready cDNA were synthesized following the manufacturer’s instructions, primed by oligo (dT) primer and the SMART II A oligonucleotide using the SMARTer RACE cDNA amplification kit (Takara Bio.).

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China). .. The amplified products were purified from 1.5% agarose gel using the Wizard SV Gel and PCR Clean-Up System (Promega, WI, USA).

Article Title: Molecular characterization and phylogenetic analysis of a dengue virus serotype 3 isolated from a Chinese traveler returned from Laos
Article Snippet: Paragraph title: Genome amplification and sequencing ... The RT reaction was conducted under the following conditions: 25 °C for 5 min, followed by 1 h at 42 °C, then 72 °C for 15 min. Next, the PCR was carried out in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTP mixture (2.5 mM), 2 μM of forward and backward primers, and one unit of high fidelity La Taq DNA polymerase (TaKaRa, Dalian, China).

Article Title: Identification and Characterization of the Gene CYP340W1 from Plutella xylostella and Its Possible Involvement in Resistance to Abamectin
Article Snippet: Rapid amplification of cDNA ends (RACE) with SMARTer RACE cDNA Amplification (Clontech, Palo Alto, CA, USA) kits was used to obtain the full-length cDNA of CYP340W1 following the manufacturer’s instructions. .. The PCR reactions (25 μL total volume) contained 15.5 μL of double-distilled H2 O (ddH2 O), 2.5 μL of 10× LA Taq Buffer, 2 μL of dNTP Mix, 1 μL of specific primer, 1.5 μL of first-strand cDNA template, 2 μL of UPM, and 0.5 μL of Advantage2 Taq HS (TaKaRa).

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: The cDNA was subsequently used for PCR amplification with an LA PCR Kit Ver.2.1 (Takara Bio, China) according to the manufacturer's protocol. .. The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total.

Article Title: Genome-wide Identification and Expression Analysis of Amino Acid Transporters in the Whitefly, Bemisia tabaci (Gennadius)
Article Snippet: Their cDNA fragments were then amplified by designing 400- to 600-bp primer pairs which were designed with Primer Premier 5.0 (Table ). .. The reactions (total 25 μl) contained 15.25 μl of ddH2 O, 4 μl of dNTP Mix, 2.5 μl of 10×LA Taq buffer, 1 μl of the cDNA template, 1 μl of each specific primer, and 0.25 μl LA Taq polymerase (TaKaRa).

Clone Assay:

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China). .. The purified products were ligated to the T4 vector (Promega, WI, USA), transformed into E. coli, and positive clones were selected for Sanger sequencing (BGI, Shenzhen).

Article Title: Genome-wide Identification and Expression Analysis of Amino Acid Transporters in the Whitefly, Bemisia tabaci (Gennadius)
Article Snippet: Paragraph title: Gene fragments cloning ... The reactions (total 25 μl) contained 15.25 μl of ddH2 O, 4 μl of dNTP Mix, 2.5 μl of 10×LA Taq buffer, 1 μl of the cDNA template, 1 μl of each specific primer, and 0.25 μl LA Taq polymerase (TaKaRa).

Electrophoresis:

Article Title: Loss of genes related to Nucleotide Excision Repair (NER) and implications for reductive genome evolution in symbionts of deep-sea vesicomyid clams
Article Snippet: For the PCR, 25 μl of the reaction mixture contained 80 ng of template DNA, 2.5 μl of 10× LA Taq buffer (TaKaRa), 2.5 μl of dNTP mix (TaKaRa), 5 μl each of the 1 pmol/μl forward and reverse primer solutions, 0.13 μl of LA Taq polymerase solution (TaKaRa) and 8.87 μl of pure water was used. .. The gene fragments were amplified with an ABI 9700 Thermal cycler using a protocol; 96°C for 2 min and 30 cycles of 96°C for 20 s, 56°C for 15 s and 72°C for 2 or 5 or 7 min, followed by extension at 72°C for 10 min. After amplification, 2 μl of the reaction product mixture was applied to 1% agarose electrophoresis gel and stained with an ethidium bromide solution to visualize the reaction products.

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total. .. The amplicons were separated by electrophoresis in a 1% agarose gel and the gene fragments were recoverd with a Gel Extraction Kit (Qiagen, Germany) according to the manufacturer's protocol.

Incubation:

Article Title: Ancient Occasional Host Switching of Maternally Transmitted Bacterial Symbionts of Chemosynthetic Vesicomyid Clams
Article Snippet: The mitochondrial or symbiont gene fragments were amplified in 25-µL PCR reactions that contained ∼100 ng DNA template, 1× LA-Taq buffer (Mg2+ plus; TaKaRa Bio, Inc., Shiga, Japan), 0.25 mM each deoxynucleotide (dNTP) solution, 0.2 µM each primer, and 0.625 U LA-Taq polymerase (TaKaRa Bio, Inc.). .. The PCR reaction mixtures were incubated under the conditions described in table 2 after the initial denaturation step (96 °C for 2 min), using a TaKara TP600 Thermal cycler (TaKaRa Bio, Inc.).

Infection:

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: 5′ / 3′ rapid amplification of cDNA ends (RACE) The viral RNA was extracted from infected cell culture supernatant using the TRIzol reagent (Invitrogen, CA, USA). .. The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China).

Touchdown PCR:

Article Title: Identification and Characterization of the Gene CYP340W1 from Plutella xylostella and Its Possible Involvement in Resistance to Abamectin
Article Snippet: The PCR reactions (25 μL total volume) contained 15.5 μL of double-distilled H2 O (ddH2 O), 2.5 μL of 10× LA Taq Buffer, 2 μL of dNTP Mix, 1 μL of specific primer, 1.5 μL of first-strand cDNA template, 2 μL of UPM, and 0.5 μL of Advantage2 Taq HS (TaKaRa). .. The touchdown PCR reaction conditions were: five cycles of 94 °C for 30 s and 72 °C for 3 min; followed by five cycles of 94 °C for 30 s, 68 °C for 30 s, and 72 °C for 3 min; and finally 25 cycles of 94 °C for 30 s, 66 °C for 30 s, and 72 °C for 3 min.

Transformation Assay:

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China). .. The purified products were ligated to the T4 vector (Promega, WI, USA), transformed into E. coli, and positive clones were selected for Sanger sequencing (BGI, Shenzhen).

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total. .. The recovered target gene product was ligated into a pMD19-T vector (Takara Bio) and used to transformed competent E. coli Top10 cells (Promega, USA).

Synthesized:

Article Title: Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera
Article Snippet: The last cycle was followed by final extension at 72°C for 10 min. Each 50 μL PCR reaction contained 2 μL of cDNA template, 5 μL of 10× LA Taq buffer (Mg2+ Free), 4 μL of MgCl2 (25 mM), 4 μL of dNTP mixture (2.5 mM/each), 1 μL of forward and 1 μL of reverse primers (10 μM), 0.5 μL of LA Taq polymerase (Takara Bio.) (5 U/μL) and 32.5 μL of double distilled H2 O. .. The 5’- and 3’-RACE Ready cDNA were synthesized following the manufacturer’s instructions, primed by oligo (dT) primer and the SMART II A oligonucleotide using the SMARTer RACE cDNA amplification kit (Takara Bio.).

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: Since the total RNA was extracted from a non-eukaryotic organism and lacked a polyadenylated tail, the 5′-first-strand cDNA was synthesized with random primers and the 3′-first-strand cDNA was synthesized after adding a poly(A) tail using Poly(A) Polymerase (TaKaRa, Dalian, China) according to the instruction of SMARTer 5′/3′ RACE kit (Clontech, CA, USA). .. The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China).

Article Title: Identification and Characterization of the Gene CYP340W1 from Plutella xylostella and Its Possible Involvement in Resistance to Abamectin
Article Snippet: First-strand cDNA was synthesized with the PrimeScript II 1st strand cDNA synthesis kit with oligo dT primer (Takara Biotechnology, Dalian, China). .. The PCR reactions (25 μL total volume) contained 15.5 μL of double-distilled H2 O (ddH2 O), 2.5 μL of 10× LA Taq Buffer, 2 μL of dNTP Mix, 1 μL of specific primer, 1.5 μL of first-strand cDNA template, 2 μL of UPM, and 0.5 μL of Advantage2 Taq HS (TaKaRa).

Cell Culture:

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: 5′ / 3′ rapid amplification of cDNA ends (RACE) The viral RNA was extracted from infected cell culture supernatant using the TRIzol reagent (Invitrogen, CA, USA). .. The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China).

Polymerase Chain Reaction:

Article Title: De Novo Assembly of Coding Sequences of the Mangrove Palm (Nypa fruticans) Using RNA-Seq and Discovery of Whole-Genome Duplications in the Ancestor of Palms
Article Snippet: PCR were set up in a 30μl reaction mixture in PCR tubes. .. Each reaction mixture contained 0.4 μM each forward and reverse primer, 0.4 mM each dNTP, 1.5 U LA Taq DNA polymerase and 3μl 10x LA Taq buffer (TaKaRa, Japan) and 50–100 ng of genomic DNA as a template.

Article Title: Identification of medium-sized genomic deletions with low coverage, mate-paired restricted tags
Article Snippet: .. PCR reactions included ~5 ng of genomic DNA, 2 μL of forward primer (10 μM), 2 μL of reverse primer (10 μM), 5 μL of 10× LA Taq Buffer (Takara ), 8 μL of dNTP (2.5 mM), 2 μL of LA Taq polymerase (Takara ) and nuclease-free water to reach a volume of 50 μL. .. Touch-down PCR was used to amplify the products.

Article Title: Ancient Occasional Host Switching of Maternally Transmitted Bacterial Symbionts of Chemosynthetic Vesicomyid Clams
Article Snippet: .. The mitochondrial or symbiont gene fragments were amplified in 25-µL PCR reactions that contained ∼100 ng DNA template, 1× LA-Taq buffer (Mg2+ plus; TaKaRa Bio, Inc., Shiga, Japan), 0.25 mM each deoxynucleotide (dNTP) solution, 0.2 µM each primer, and 0.625 U LA-Taq polymerase (TaKaRa Bio, Inc.). .. The PCR reaction mixtures were incubated under the conditions described in table 2 after the initial denaturation step (96 °C for 2 min), using a TaKara TP600 Thermal cycler (TaKaRa Bio, Inc.).

Article Title: Application of next-generation sequencing to characterize novel mutations in clarithromycin-susceptible Helicobacter pylori strains with A2143G of 23S rRNA gene
Article Snippet: .. The PCR reaction was performed in 25-µl reactions containing 2.5 µl of 10× LA Taq Buffer, 4 µl of dNTP mixture (2.5 mM each), 0.5 µl each 10 µM primer, 2 µl template DNA and 0.25 µl of TaKaRa LA Taq™ (5 units/µl). ..

Article Title: Loss of genes related to Nucleotide Excision Repair (NER) and implications for reductive genome evolution in symbionts of deep-sea vesicomyid clams
Article Snippet: .. For the PCR, 25 μl of the reaction mixture contained 80 ng of template DNA, 2.5 μl of 10× LA Taq buffer (TaKaRa), 2.5 μl of dNTP mix (TaKaRa), 5 μl each of the 1 pmol/μl forward and reverse primer solutions, 0.13 μl of LA Taq polymerase solution (TaKaRa) and 8.87 μl of pure water was used. .. The gene fragments were amplified with an ABI 9700 Thermal cycler using a protocol; 96°C for 2 min and 30 cycles of 96°C for 20 s, 56°C for 15 s and 72°C for 2 or 5 or 7 min, followed by extension at 72°C for 10 min. After amplification, 2 μl of the reaction product mixture was applied to 1% agarose electrophoresis gel and stained with an ethidium bromide solution to visualize the reaction products.

Article Title: Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera
Article Snippet: .. The last cycle was followed by final extension at 72°C for 10 min. Each 50 μL PCR reaction contained 2 μL of cDNA template, 5 μL of 10× LA Taq buffer (Mg2+ Free), 4 μL of MgCl2 (25 mM), 4 μL of dNTP mixture (2.5 mM/each), 1 μL of forward and 1 μL of reverse primers (10 μM), 0.5 μL of LA Taq polymerase (Takara Bio.) (5 U/μL) and 32.5 μL of double distilled H2 O. .. The 5’- and 3’-RACE Ready cDNA were synthesized following the manufacturer’s instructions, primed by oligo (dT) primer and the SMART II A oligonucleotide using the SMARTer RACE cDNA amplification kit (Takara Bio.).

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: .. The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China). ..

Article Title: Molecular characterization and phylogenetic analysis of a dengue virus serotype 3 isolated from a Chinese traveler returned from Laos
Article Snippet: .. The RT reaction was conducted under the following conditions: 25 °C for 5 min, followed by 1 h at 42 °C, then 72 °C for 15 min. Next, the PCR was carried out in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTP mixture (2.5 mM), 2 μM of forward and backward primers, and one unit of high fidelity La Taq DNA polymerase (TaKaRa, Dalian, China). .. PCR amplification was performed under the following parameters [ ]: 94 °C for 5 min, followed by 35 cycles of denaturation at 94 °C for 30 s, annealing at 55 °C for 30 s, extension at 72 °C for 2 min and final extension at 72 °C for 5 min.

Article Title: Identification and Characterization of the Gene CYP340W1 from Plutella xylostella and Its Possible Involvement in Resistance to Abamectin
Article Snippet: .. The PCR reactions (25 μL total volume) contained 15.5 μL of double-distilled H2 O (ddH2 O), 2.5 μL of 10× LA Taq Buffer, 2 μL of dNTP Mix, 1 μL of specific primer, 1.5 μL of first-strand cDNA template, 2 μL of UPM, and 0.5 μL of Advantage2 Taq HS (TaKaRa). .. The touchdown PCR reaction conditions were: five cycles of 94 °C for 30 s and 72 °C for 3 min; followed by five cycles of 94 °C for 30 s, 68 °C for 30 s, and 72 °C for 3 min; and finally 25 cycles of 94 °C for 30 s, 66 °C for 30 s, and 72 °C for 3 min.

Article Title: Identification and Characterization of the Gene CYP340W1 from Plutella xylostella and Its Possible Involvement in Resistance to Abamectin
Article Snippet: .. The PCR reactions (25 μL total volume) contained, 2.5 μL of 10× LA Taq Buffer, 2 μL of dNTP Mix, 1 μL of first-strand cDNA template, 1 μL of specific primer, 0.25 μL LA Taq HS polymerase (TaKaRa), and 18.25 μL of double-distilled H2 O (ddH2 O). .. All PCR reactions were carried out with a S1000 PCR Thermal Cycler (Bio-Rad, Shanghai, China).

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: .. The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total. ..

Article Title: Genome-wide Identification and Expression Analysis of Amino Acid Transporters in the Whitefly, Bemisia tabaci (Gennadius)
Article Snippet: The reactions (total 25 μl) contained 15.25 μl of ddH2 O, 4 μl of dNTP Mix, 2.5 μl of 10×LA Taq buffer, 1 μl of the cDNA template, 1 μl of each specific primer, and 0.25 μl LA Taq polymerase (TaKaRa). .. The PCR reactions were then performed using a S1000 PCR system (BioRad).

DNA Sequencing:

Article Title: Molecular characterization and phylogenetic analysis of a dengue virus serotype 3 isolated from a Chinese traveler returned from Laos
Article Snippet: The RT reaction was conducted under the following conditions: 25 °C for 5 min, followed by 1 h at 42 °C, then 72 °C for 15 min. Next, the PCR was carried out in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTP mixture (2.5 mM), 2 μM of forward and backward primers, and one unit of high fidelity La Taq DNA polymerase (TaKaRa, Dalian, China). .. Finally, the PCR products were used as templates for bi-directional DNA sequencing by the Sanger dideoxy sequencing method (Invitrogen Ltd., Shanghai).

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: Paragraph title: Preparation of viral RNA, reverse transcription PCR, and DNA sequencing ... The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total.

Sequencing:

Article Title: De Novo Assembly of Coding Sequences of the Mangrove Palm (Nypa fruticans) Using RNA-Seq and Discovery of Whole-Genome Duplications in the Ancestor of Palms
Article Snippet: Each reaction mixture contained 0.4 μM each forward and reverse primer, 0.4 mM each dNTP, 1.5 U LA Taq DNA polymerase and 3μl 10x LA Taq buffer (TaKaRa, Japan) and 50–100 ng of genomic DNA as a template. .. Twenty-five of them successfully got the PCR products and were sequenced by Sanger sequencing.

Article Title: Identification of medium-sized genomic deletions with low coverage, mate-paired restricted tags
Article Snippet: Paragraph title: Validation by PCR amplification and clone sequencing ... PCR reactions included ~5 ng of genomic DNA, 2 μL of forward primer (10 μM), 2 μL of reverse primer (10 μM), 5 μL of 10× LA Taq Buffer (Takara ), 8 μL of dNTP (2.5 mM), 2 μL of LA Taq polymerase (Takara ) and nuclease-free water to reach a volume of 50 μL.

Article Title: Ancient Occasional Host Switching of Maternally Transmitted Bacterial Symbionts of Chemosynthetic Vesicomyid Clams
Article Snippet: Paragraph title: Polymerase Chain Reaction (PCR) Amplification and Sequencing ... The mitochondrial or symbiont gene fragments were amplified in 25-µL PCR reactions that contained ∼100 ng DNA template, 1× LA-Taq buffer (Mg2+ plus; TaKaRa Bio, Inc., Shiga, Japan), 0.25 mM each deoxynucleotide (dNTP) solution, 0.2 µM each primer, and 0.625 U LA-Taq polymerase (TaKaRa Bio, Inc.).

Article Title: Integrating nested PCR with high-throughput sequencing to characterize mutations of HBV genome in low viral load samples
Article Snippet: 2.3 Primers and amplification The complete HBV genome was amplified using nested primers (Table ) and validated with Sanger sequencing using specific primers. .. The reaction contained 2 μL of template DNA, 0.5 μL of each 10 μM primer, 2.5 μL of 10× LA Taq Buffer (MgCl2 -free), 4 μL of dNTP mixture (2.5 mM each), 2.5 μL of 25 mM MgCl2 , and 0.25 μL of TaKaRa LA Taq (5 units/μL).

Article Title: Application of next-generation sequencing to characterize novel mutations in clarithromycin-susceptible Helicobacter pylori strains with A2143G of 23S rRNA gene
Article Snippet: Paragraph title: Direct sequencing characterized the mutations of 23S rRNA ... The PCR reaction was performed in 25-µl reactions containing 2.5 µl of 10× LA Taq Buffer, 4 µl of dNTP mixture (2.5 mM each), 0.5 µl each 10 µM primer, 2 µl template DNA and 0.25 µl of TaKaRa LA Taq™ (5 units/µl).

Article Title: Loss of genes related to Nucleotide Excision Repair (NER) and implications for reductive genome evolution in symbionts of deep-sea vesicomyid clams
Article Snippet: Paragraph title: Amplification and sequencing of NER genes ... For the PCR, 25 μl of the reaction mixture contained 80 ng of template DNA, 2.5 μl of 10× LA Taq buffer (TaKaRa), 2.5 μl of dNTP mix (TaKaRa), 5 μl each of the 1 pmol/μl forward and reverse primer solutions, 0.13 μl of LA Taq polymerase solution (TaKaRa) and 8.87 μl of pure water was used.

Article Title: Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera
Article Snippet: The final cDNA sequence was authenticated using the primers listed in Table . .. The last cycle was followed by final extension at 72°C for 10 min. Each 50 μL PCR reaction contained 2 μL of cDNA template, 5 μL of 10× LA Taq buffer (Mg2+ Free), 4 μL of MgCl2 (25 mM), 4 μL of dNTP mixture (2.5 mM/each), 1 μL of forward and 1 μL of reverse primers (10 μM), 0.5 μL of LA Taq polymerase (Takara Bio.) (5 U/μL) and 32.5 μL of double distilled H2 O.

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China). .. The purified products were ligated to the T4 vector (Promega, WI, USA), transformed into E. coli, and positive clones were selected for Sanger sequencing (BGI, Shenzhen).

Article Title: Molecular characterization and phylogenetic analysis of a dengue virus serotype 3 isolated from a Chinese traveler returned from Laos
Article Snippet: Paragraph title: Genome amplification and sequencing ... The RT reaction was conducted under the following conditions: 25 °C for 5 min, followed by 1 h at 42 °C, then 72 °C for 15 min. Next, the PCR was carried out in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTP mixture (2.5 mM), 2 μM of forward and backward primers, and one unit of high fidelity La Taq DNA polymerase (TaKaRa, Dalian, China).

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: The forward primer sequence was 5'-CGGGATCC ATGTTGGGGAAGTGCT-3' (containing a BamH I site in the 5' terminal) and the reverse primer sequence was 5'-CGGAATTC CTAGAGACGACCCCATT-3' (containing an EcoR I site in 5' terminal). .. The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total.

Imaging:

Article Title: Molecular characterization and phylogenetic analysis of a dengue virus serotype 3 isolated from a Chinese traveler returned from Laos
Article Snippet: The RT reaction was conducted under the following conditions: 25 °C for 5 min, followed by 1 h at 42 °C, then 72 °C for 15 min. Next, the PCR was carried out in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTP mixture (2.5 mM), 2 μM of forward and backward primers, and one unit of high fidelity La Taq DNA polymerase (TaKaRa, Dalian, China). .. The PCR results were analyzed by 1% agarose gel electrophoresis and visualized by a biospectrum® 815 imaging system (UVP, USA).

Molecular Cloning:

Article Title: Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera
Article Snippet: Paragraph title: Molecular cloning ... The last cycle was followed by final extension at 72°C for 10 min. Each 50 μL PCR reaction contained 2 μL of cDNA template, 5 μL of 10× LA Taq buffer (Mg2+ Free), 4 μL of MgCl2 (25 mM), 4 μL of dNTP mixture (2.5 mM/each), 1 μL of forward and 1 μL of reverse primers (10 μM), 0.5 μL of LA Taq polymerase (Takara Bio.) (5 U/μL) and 32.5 μL of double distilled H2 O.

Article Title: Identification and Characterization of the Gene CYP340W1 from Plutella xylostella and Its Possible Involvement in Resistance to Abamectin
Article Snippet: Paragraph title: 4.3. Molecular Cloning of CYP340W1 ... The PCR reactions (25 μL total volume) contained 15.5 μL of double-distilled H2 O (ddH2 O), 2.5 μL of 10× LA Taq Buffer, 2 μL of dNTP Mix, 1 μL of specific primer, 1.5 μL of first-strand cDNA template, 2 μL of UPM, and 0.5 μL of Advantage2 Taq HS (TaKaRa).

Size-exclusion Chromatography:

Article Title: De Novo Assembly of Coding Sequences of the Mangrove Palm (Nypa fruticans) Using RNA-Seq and Discovery of Whole-Genome Duplications in the Ancestor of Palms
Article Snippet: Each reaction mixture contained 0.4 μM each forward and reverse primer, 0.4 mM each dNTP, 1.5 U LA Taq DNA polymerase and 3μl 10x LA Taq buffer (TaKaRa, Japan) and 50–100 ng of genomic DNA as a template. .. PCR amplification was carried out using a Dyad Disciple thermal Cycler (Bio-Rad, Beijing, USA) with the following cycling conditions: pre-denaturation at 94°C for 5 min followed by 33 cycles of 94°C for 30 sec, 57°C for 45 sec and 72°C for 1 min 30 sec, and finally, 10 min at 72°C for extension.

Article Title: Molecular cloning and RNA interference-mediated functional characterization of a Halloween gene spook in the white-backed planthopper Sogatella furcifera
Article Snippet: Thermal cycling conditions were 94°C for 5 min, followed by 35 cycles of 94°C for 30 sec, 55°C for 45 sec and 72°C for 3 min. .. The last cycle was followed by final extension at 72°C for 10 min. Each 50 μL PCR reaction contained 2 μL of cDNA template, 5 μL of 10× LA Taq buffer (Mg2+ Free), 4 μL of MgCl2 (25 mM), 4 μL of dNTP mixture (2.5 mM/each), 1 μL of forward and 1 μL of reverse primers (10 μM), 0.5 μL of LA Taq polymerase (Takara Bio.) (5 U/μL) and 32.5 μL of double distilled H2 O.

Purification:

Article Title: Integrating nested PCR with high-throughput sequencing to characterize mutations of HBV genome in low viral load samples
Article Snippet: The reaction contained 2 μL of template DNA, 0.5 μL of each 10 μM primer, 2.5 μL of 10× LA Taq Buffer (MgCl2 -free), 4 μL of dNTP mixture (2.5 mM each), 2.5 μL of 25 mM MgCl2 , and 0.25 μL of TaKaRa LA Taq (5 units/μL). .. PCR products were analyzed in 1.5% agarose gel electrophoresis and cleaned up with Qiagen PCR Purification Kit (Qiagen, Valencia, CA).

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China). .. The amplified products were purified from 1.5% agarose gel using the Wizard SV Gel and PCR Clean-Up System (Promega, WI, USA).

Reverse Transcription Polymerase Chain Reaction:

Article Title: Loss of genes related to Nucleotide Excision Repair (NER) and implications for reductive genome evolution in symbionts of deep-sea vesicomyid clams
Article Snippet: The specific RT-PCR primer sets for NER genes were designed from aligned sequences of Vok and the C . pacifica symbiont (Cpac_S). .. For the PCR, 25 μl of the reaction mixture contained 80 ng of template DNA, 2.5 μl of 10× LA Taq buffer (TaKaRa), 2.5 μl of dNTP mix (TaKaRa), 5 μl each of the 1 pmol/μl forward and reverse primer solutions, 0.13 μl of LA Taq polymerase solution (TaKaRa) and 8.87 μl of pure water was used.

Article Title: Molecular characterization and phylogenetic analysis of a dengue virus serotype 3 isolated from a Chinese traveler returned from Laos
Article Snippet: Genome amplification and sequencing A total of nine overlapping sub-genomic fragments that spanned the complete genomic region were amplified using nine pairs of primers (Additional file : Table S2) [ ] by RT-PCR. .. The RT reaction was conducted under the following conditions: 25 °C for 5 min, followed by 1 h at 42 °C, then 72 °C for 15 min. Next, the PCR was carried out in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTP mixture (2.5 mM), 2 μM of forward and backward primers, and one unit of high fidelity La Taq DNA polymerase (TaKaRa, Dalian, China).

Staining:

Article Title: Ancient Occasional Host Switching of Maternally Transmitted Bacterial Symbionts of Chemosynthetic Vesicomyid Clams
Article Snippet: The mitochondrial or symbiont gene fragments were amplified in 25-µL PCR reactions that contained ∼100 ng DNA template, 1× LA-Taq buffer (Mg2+ plus; TaKaRa Bio, Inc., Shiga, Japan), 0.25 mM each deoxynucleotide (dNTP) solution, 0.2 µM each primer, and 0.625 U LA-Taq polymerase (TaKaRa Bio, Inc.). .. To check the size of the PCR-amplified fragments, 2.0-µL aliquots of the PCR reaction mixtures were electrophoresed on 0.6–1.0% agarose gels and stained with 0.06% ethidium bromide.

Article Title: Loss of genes related to Nucleotide Excision Repair (NER) and implications for reductive genome evolution in symbionts of deep-sea vesicomyid clams
Article Snippet: For the PCR, 25 μl of the reaction mixture contained 80 ng of template DNA, 2.5 μl of 10× LA Taq buffer (TaKaRa), 2.5 μl of dNTP mix (TaKaRa), 5 μl each of the 1 pmol/μl forward and reverse primer solutions, 0.13 μl of LA Taq polymerase solution (TaKaRa) and 8.87 μl of pure water was used. .. The gene fragments were amplified with an ABI 9700 Thermal cycler using a protocol; 96°C for 2 min and 30 cycles of 96°C for 20 s, 56°C for 15 s and 72°C for 2 or 5 or 7 min, followed by extension at 72°C for 10 min. After amplification, 2 μl of the reaction product mixture was applied to 1% agarose electrophoresis gel and stained with an ethidium bromide solution to visualize the reaction products.

Nested PCR:

Article Title: Integrating nested PCR with high-throughput sequencing to characterize mutations of HBV genome in low viral load samples
Article Snippet: [ , ] The HBV genome was divided into 2 fragments (Fig. ) and both rounds of nested PCR were performed in 25 μL reaction system. .. The reaction contained 2 μL of template DNA, 0.5 μL of each 10 μM primer, 2.5 μL of 10× LA Taq Buffer (MgCl2 -free), 4 μL of dNTP mixture (2.5 mM each), 2.5 μL of 25 mM MgCl2 , and 0.25 μL of TaKaRa LA Taq (5 units/μL).

Rapid Amplification of cDNA Ends:

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: Paragraph title: 5′ / 3′ rapid amplification of cDNA ends (RACE) ... The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China).

Article Title: Identification and Characterization of the Gene CYP340W1 from Plutella xylostella and Its Possible Involvement in Resistance to Abamectin
Article Snippet: Rapid amplification of cDNA ends (RACE) with SMARTer RACE cDNA Amplification (Clontech, Palo Alto, CA, USA) kits was used to obtain the full-length cDNA of CYP340W1 following the manufacturer’s instructions. .. The PCR reactions (25 μL total volume) contained 15.5 μL of double-distilled H2 O (ddH2 O), 2.5 μL of 10× LA Taq Buffer, 2 μL of dNTP Mix, 1 μL of specific primer, 1.5 μL of first-strand cDNA template, 2 μL of UPM, and 0.5 μL of Advantage2 Taq HS (TaKaRa).

Plasmid Preparation:

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China). .. The purified products were ligated to the T4 vector (Promega, WI, USA), transformed into E. coli, and positive clones were selected for Sanger sequencing (BGI, Shenzhen).

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total. .. The recovered target gene product was ligated into a pMD19-T vector (Takara Bio) and used to transformed competent E. coli Top10 cells (Promega, USA).

Software:

Article Title: De Novo Assembly of Coding Sequences of the Mangrove Palm (Nypa fruticans) Using RNA-Seq and Discovery of Whole-Genome Duplications in the Ancestor of Palms
Article Snippet: To validate the predicted SSRs, we used Primer3 software [ ] to design primers around the SSRs. .. Each reaction mixture contained 0.4 μM each forward and reverse primer, 0.4 mM each dNTP, 1.5 U LA Taq DNA polymerase and 3μl 10x LA Taq buffer (TaKaRa, Japan) and 50–100 ng of genomic DNA as a template.

Article Title: Identification of medium-sized genomic deletions with low coverage, mate-paired restricted tags
Article Snippet: Validation by PCR amplification and clone sequencing PCR primers were designed based on the ditags using Primer Premier 5 software (Additional file : Table S3). .. PCR reactions included ~5 ng of genomic DNA, 2 μL of forward primer (10 μM), 2 μL of reverse primer (10 μM), 5 μL of 10× LA Taq Buffer (Takara ), 8 μL of dNTP (2.5 mM), 2 μL of LA Taq polymerase (Takara ) and nuclease-free water to reach a volume of 50 μL.

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: To obtain the complete ORF5 gene of the PRRSV Guizhou strain, one pair of specific primers was designed using Primer Premier 5.0 software (PREMIER Biosoft, Canada) according to the ORF5 gene sequence of PRRSV Guizhou-1 strain (GenBank No. EU259060), CH-1a strain (GenBank No. AY032626), and VR-2332 strain (GenBank No. PRU87392) published in GenBank. .. The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total.

Agarose Gel Electrophoresis:

Article Title: Integrating nested PCR with high-throughput sequencing to characterize mutations of HBV genome in low viral load samples
Article Snippet: The reaction contained 2 μL of template DNA, 0.5 μL of each 10 μM primer, 2.5 μL of 10× LA Taq Buffer (MgCl2 -free), 4 μL of dNTP mixture (2.5 mM each), 2.5 μL of 25 mM MgCl2 , and 0.25 μL of TaKaRa LA Taq (5 units/μL). .. PCR products were analyzed in 1.5% agarose gel electrophoresis and cleaned up with Qiagen PCR Purification Kit (Qiagen, Valencia, CA).

Article Title: Application of next-generation sequencing to characterize novel mutations in clarithromycin-susceptible Helicobacter pylori strains with A2143G of 23S rRNA gene
Article Snippet: The PCR reaction was performed in 25-µl reactions containing 2.5 µl of 10× LA Taq Buffer, 4 µl of dNTP mixture (2.5 mM each), 0.5 µl each 10 µM primer, 2 µl template DNA and 0.25 µl of TaKaRa LA Taq™ (5 units/µl). .. 1.2% agarose gel electrophoresis was utilized for verifying the PCR products size at 360 bp.

Article Title: Full-length genome and molecular characterization of dengue virus serotype 2 isolated from an imported patient from Myanmar
Article Snippet: The second round PCRs were performed in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTPs (2.5 mM), 5 μL of 10 × Universal Primer Mix, 1 μL of 5′/3′ Gene-Specific Primer, 2.5 μL of diluted 5′/3′ RACE PCR products and 1 unit of high fidelity La Taq DAN polymerase (TaKaRa, Dalian, China). .. The amplified products were purified from 1.5% agarose gel using the Wizard SV Gel and PCR Clean-Up System (Promega, WI, USA).

Article Title: Molecular characterization and phylogenetic analysis of a dengue virus serotype 3 isolated from a Chinese traveler returned from Laos
Article Snippet: The RT reaction was conducted under the following conditions: 25 °C for 5 min, followed by 1 h at 42 °C, then 72 °C for 15 min. Next, the PCR was carried out in a reaction mixture of 50 μL containing 5 μL of 10 × La Taq Buffer (Mg2+ Plus), 8 μL of dNTP mixture (2.5 mM), 2 μM of forward and backward primers, and one unit of high fidelity La Taq DNA polymerase (TaKaRa, Dalian, China). .. The PCR results were analyzed by 1% agarose gel electrophoresis and visualized by a biospectrum® 815 imaging system (UVP, USA).

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total. .. The amplicons were separated by electrophoresis in a 1% agarose gel and the gene fragments were recoverd with a Gel Extraction Kit (Qiagen, Germany) according to the manufacturer's protocol.

Ethanol Precipitation:

Article Title: Loss of genes related to Nucleotide Excision Repair (NER) and implications for reductive genome evolution in symbionts of deep-sea vesicomyid clams
Article Snippet: For the PCR, 25 μl of the reaction mixture contained 80 ng of template DNA, 2.5 μl of 10× LA Taq buffer (TaKaRa), 2.5 μl of dNTP mix (TaKaRa), 5 μl each of the 1 pmol/μl forward and reverse primer solutions, 0.13 μl of LA Taq polymerase solution (TaKaRa) and 8.87 μl of pure water was used. .. After ethanol precipitation, the nucleotide sequences of the amplified DNA were determined using a Big Dye Terminator v3.1 Cycle Sequencing Kit (Applied Biosystems) with an ABI PRIZM 3130xl Genetic Analyzer and ABI 3730xl Genetic Analyzer (Applied Biosystems) according to the manufacturer’s instruction.

Gel Extraction:

Article Title: Positive effects of porcine IL-2 and IL-4 on virus-specific immune responses induced by the porcine reproductive and respiratory syndrome virus (PRRSV) ORF5 DNA vaccine in swine
Article Snippet: The PCR reaction system as follows: 10× LA Taq Buffer 5 µL (Takara Bio), 2.5 mM dNTP Mixture 4 µL (Takara Bio), cDNA 2 µL, 2 µL each of forward and reverse primer (20 pmol/µL), LA Taq polymerase 1 µL (5 U/µL) (Takara Bio), adding ddH2O to 50 µL in total. .. The amplicons were separated by electrophoresis in a 1% agarose gel and the gene fragments were recoverd with a Gel Extraction Kit (Qiagen, Germany) according to the manufacturer's protocol.

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