Quick taq hs dyemix

Manufactured by Toyobo

Sourced in Japan, United States

Quick Taq HS DyeMix is a ready-to-use, high-speed PCR master mix that contains all the necessary components for efficient DNA amplification, including a hot-start DNA polymerase, nucleotides, and reaction buffer.

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Lab products found in correlation

Trizol, by Thermo Fisher Scientific (18 mentions) T100 thermal cycler, by Bio-Rad (7 mentions) Primescript rt reagent kit, by Takara Bio (7 mentions) Revertra ace qpcr rt master mix, by Toyobo (6 mentions) Rneasy mini kit, by Qiagen (3 mentions) Trizol reagent, by Thermo Fisher Scientific (3 mentions) Topo ta cloning kit, by Thermo Fisher Scientific (2 mentions) Dig rna labeling mix, by Roche (2 mentions) Revertra ace, by Toyobo (2 mentions) Wizard sv gel and pcr clean up system, by Promega (2 mentions) Primescript 1st strand cdna synthesis kit, by Takara Bio (2 mentions) Ultraviolet light, by Bio-Rad (2 mentions) Gdna remover, by Toyobo (2 mentions) Nucleospin rna plus, by Macherey-Nagel (2 mentions) Rneasy plant mini kit, by Qiagen (2 mentions) Expin pcr sv kit, by GeneAll (2 mentions) Revertra ace α, by Toyobo (2 mentions) Dna engine, by Bio-Rad (2 mentions) Fastgene gel pcr extraction kit, by Nippon Genetics (2 mentions) Dneasy blood and tissue kit, by Qiagen (2 mentions)

Spelling variants of this product

Quick TaqTM HS DyeMix (5 citations) Quick Taq HS (2 citations) Quick Taq HS DyeMix DNA polymerase (2 citations)

77 protocols using quick taq hs dyemix

Viral RNA Detection via RT-PCR

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Toward viral RNA detection by RT-PCR, the ssRNA-enriched fractions described above were used for cDNA synthesis by M-MLV Reverse Transcriptase (Invitrogen, Thermo Fisher Scientific Inc.) with the random primer [hexadeoxyribonucleotide mixture; pd (N)₆] (TaKaRa Bio Inc.) according to the manufacturer’s instruction in a half scale. The cDNA was used as a template for PCR by Quick Taq HS DyeMix (TOYOBO Co., Ltd.) on a 10-μl scale. PCR-based genotyping of R. necatrix was also performed by Quick Taq HS DyeMix (TOYOBO Co., Ltd.) on a 10-μl scale with the host genomic DNA template. Primers are listed in Table S5.
The RT-PCR and genomic PCR products were electrophoresed in 1% (wt/vol) agarose in 0.5× TAE. The nucleic acids were visualized with ethidium bromide (EtBr) by postgel staining. GeneRuler 1-kb DNA ladder (Thermo Fischer Scientific, Inc.) (described as “M-dsDNA” in the figures) was constantly used as a molecular size marker for nucleic acids.

Sato Y., Hisano S., López-Herrera C.J., Kondo H, & Suzuki N. (2022). Three-Layered Complex Interactions among Capsidless (+)ssRNA Yadokariviruses, dsRNA Viruses, and a Fungus. mBio, 13(5), e01685-22.

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PCR Amplification of Artificial Chromosome DNA

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To assess the capability of recovering the DNA information stored in an artificial chromosome, a polymerase chain reaction (PCR) was conducted to amplify a specific DNA segment of the linear plasmid DNA. The specific 310 bp segment was amplified using the sense primer 5′-ACG GAG ACT GGA GTC GAA GAG G-3′ and the antisense primer 5′-GTA GGG CAA CTA GTG CAT CTC CC-3′. Each reaction mixture (50 μL) contained 50 ng of DNA, 10 pmol of each primer, and 25 μL of 2× Quick Taq HS DyeMix (Toyobo, Japan). PCR amplification (30 cycles) was performed with a T100 Thermal Cycler (Bio-Rad, USA). Each cycle included an initial denaturation step at 94 °C for 2 min, a denaturation step at 94 °C for 30 s, a primer annealing step at 58 °C for 30 s, and a extension step at 68 °C for 1 min. The T_m of the sense primer was 65.9 °C and the T_m of the antisense primer was 66.4 °C. After the 30 cycles of amplification, the final extension step was conducted at 68 °C for 5 min. Then the samples were stored at 4 °C until use. The amplified products were investigated by agarose gel electrophoresis. The electrophoresis was conducted for 120 min at 80 V in a 2 % agarose gel using 1× TBE buffer. SYBR Safe DNA Gel Stain (Invitrogen, USA) was used for staining.

Jung Y.J., Kim H., Cheong H.K, & Lim Y.B. (2023). Magnetic control of self-assembly and disassembly in organic materials. Nature Communications, 14, 3081.

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Polymorphism Analysis of Rice Accessions

Cited 4 times

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Total DNA was extracted from young leaves of individual plants by the CTAB method, and polymorphism data of 67 SSR markers distributed among the 12 chromosomes (Supplemental Table 2) were collected. Kawasaki-Tanaka and Fukuta (2014) (link), Wunna et al. (2016) (link), Khan et al. (2017) (link) and Odjo et al. (2017) (link) also used the SSR markers, and they demonstrated that these polymorphisms could differentiate among the upland Japonica, lowland Japonica and Indica Groups. PCR was performed in a 10-μL mixture containing 1 μL sterile H₂O, a total 1.5 μL of forward and reverse primers (each 2 μM), 7.5 μL of 2× Quick Taq HS Dye Mix (Toyobo Co., Ltd. Japan) and 5 μL DNA concentrated to about 5 to 10 ng/μL. PCR amplification was performed with the following profile: 94°C for 2 min, followed by 40 cycles of 30 s at 94°C, 30 s at 55°C and 1 min at 68°C. The amplified products were separated by electrophoresis in 2% agarose gel in 1× TAE buffer at 150 V for 90 or 120 min, and DNA polymorphisms were detected by ethidium bromide staining.
The banding patterns were scored based on the presence (1) or absence (0) in each SSR markers’ allele. These polymorphism data were used to classify the accessions by Ward’s hierarchical clustering method in JMP software (v. 7.0.2; SAS Institute, Inc., Cary, NC, USA).

Muto C., Ebana K., Kawano K., Bounphanousay V., Bounphanousay C., Kanyavong K., Inthapanya P., Boualaphanh C., Sato T., Ishikawa R., Sato Y.I., Yanagihara S, & Fukuta Y. (2019). Genetic variation in rice (Oryza sativa L.) germplasm from northern Laos. Breeding Science, 69(2), 272-278.

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Evaluating Rice Trait Mutations

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Regarding hull color, pericarp color, and glutinosity, major loss-of-function mutations have been reported to be caused by the 22 bp deletion at Bh4, 14 bp deletion at Rc, and 23 bp insertion at Waxy loci [19 (link),20 (link),21 (link)]. Primer pairs were designed to detect differences in length (Table S7). They were subjected to PCR in a 20 μL mixture containing 0.2 μM of each primer and 10 μL of 2× Quick Taq HS DyeMix (TOYOBO). Amplification was carried out as follows: initial denaturation at 94 °C for 5 min, followed by 35 cycles of 30 s at 94 °C, 30 s at 55 °C, 1 min at 72 °C, and a final extension for 5 min at 72 °C. The PCR products were electrophoresed on a 4% polyacrylamide gel, and band patterns were visualized using the silver staining method [24 (link)]. Based on the band sizes, functional and non-functional alleles were estimated at the loci for hull color, pericarp color, and glutinosity.

Lim S., Onoda A., Orn C., Iwamoto H., Ishikawa R., Saito H., Sato Y, & Ishii T. (2022). Variations in Grain Traits among Local Rice Varieties Collected More Than Half-Century Ago in Indochinese Countries. Plants, 12(1), 133.

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Screening for Antimicrobial Resistance Genes

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The following 20 antimicrobial genes were investigated: cepA, cfxA, cfiA (β-lactamase), ermB, ermF, ermG, linA, mefA, msrSA [macrolide–lincomycin–streptogramin (MLS) resistant], tetM, tetQ, tetX, tetX1, tet36 (tetracycline resistant), bexA, qnrA, qnrB, qnrS (quinolone resistant), nim (metronidazole resistant), and catA (chloramphenicol resistant). For those strains found to harbor cfiA, further investigation of three insertion sequences (IS1186, IS1187, and IS942) was carried out.
Bacterial cells from the colonies on the surface of anaerobic agar plates were suspended in 0.5 mL water in 1.5 mL Eppendorf tubes and incubated at 95 °C for 8 min. The supernatants of the centrifuged suspensions (5 min, 16,000 x g) were used as template DNA and stored at −20 °C. The template DNA (5 µL) was amplified in a 45 µL-reaction mixture consisting of 25 µL of 2x Quick Taq^® HS DyeMix (Toyobo Co. Ltd, Osaka, Japan), 2 µL of each primer, and 16 µL of distilled water. Amplification was performed using a Takara PCR Thermal Cycler Dice TP 600 (Takara Bio Inc., Shiga, Japan). The PCR conditions used to detect all 20 genes and 3 insert sequences, primer sequences, and PCR parameters are listed in

Table S1

. The PCR products were examined by electrophoresis on a 2% agarose gel.

Vu H., Hayashi M., Nguyen T.N., Khong D.T., Tran H.T., Yamamoto Y, & Tanaka K. (2021). Comparison of Phenotypic and Genotypic Patterns of Antimicrobial-Resistant Bacteroides fragilis Group Isolated from Healthy Individuals in Vietnam and Japan. Infection and Drug Resistance, 14, 5313-5323.

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NPM1 Mutation Detection by PCR Assay

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The standard PCR assay was performed on 83 patient samples to detect NPM1 mutations. The PCR reaction in a final volume 20 μL contained 10 μL of 2X Quick Taq HS DyeMix (Toyobo, Japan), 0.2 μM of forward primer (NPM-F:5’-GTGTTGTGGTTCCTTAACCACAT-3’) and reverse primer (5’-CTGTTACAG AAATGAAATAAGACGGAAA-3’), approximately 100 ng of patient DNA, and ddH20 (up to 20 μL). KG-1a DNA and synthesized mutated-NPM1 ssDNA fragments were used as the wild-type control and mutated control, respectively. PCR was performed in a Mini MJ Thermal Cycler (Bio-rad, Inc., Singapore), including initial denaturation 95°C for 3 min, followed by 35 cycles of 95°C for 30 s, 60°C for 30 s and 72°C for 30 s, with a final extension at 72°C for 3 min. PCR products were obtained on 10% polyacrylamide gel electrophoresis using vertical gel electrophoresis, run at 400 volts for 150 min and visualized by a molecular imager to determine the amplicon of both wild-type and mutant products. All PCR products were confirmed by direct sequencing in both directions using fluorescent dye-terminator sequencing on ABI 3730xl DNA sequencers (Applied Biosystems^™, Thermo Fisher Scientific Inc. MA, USA).

Kongta R., Panyasit N., Jansaento W, & Duangmano S. (2022). Development of E-ice-COLD-PCR assay combined with HRM analysis for Nucleophosmin1 gene mutation detection in acute myelogenous leukemia. PLoS ONE, 17(9), e0274034.

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FLT3-ITD Mutation Detection Protocol

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A primer pair was designed to specifically amplify both of FLT3-ITD mutant and FLT3-wild type at exon 14-15 (F 5′-GCAATTTAGGTATGAAAGCCAGC-3′ and R 5′-CTTTCAG-CATTTTGACGGCAAC-3′) yielding a 300 bp wild-type PCR product. Any patient with an additional higher molecular weight band was considered to be FLT3-ITD mutant. The conventional PCR for FLT3 mutation was performed on the MyCycler Thermal Cycler machine (BIO-RAD). Positive and negative controls, as well as a blank control with distilled water, were included in each run of unknown samples. Here, 50 ng/µL of DNA samples were amplified in a total volume of 20 µL containing 0.2 µM of each primer and 10 µL of 2X Quick Taq Hs DyeMix (Toyobo, Japan). PCR was performed at 95°C for 3 minutes followed by 40 cycles at 95°C for 30 seconds, 60°C for 30 seconds, and 72°C for 30 seconds, with a final extension step at 72°C for 3 minutes. The PCR products were separated by electrophoresis through 3% agarose gel electrophoresis and the PCR product bands were viewed under UV illumination. Cases in which an additional higher molecular weight band was identified were considered FLT3-ITD-positive.

Kongta R., Thanomwongkhan K., Panyasit N., & Wang W. (2022). Rapid detection of FLT3-ITD (exon14-15) gene mutations analysis in acute myelogenous leukemia patients by High Resolution Melting analysis. Journal of Associated Medical Sciences, 55(1).

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Genetic Diversity of Japanese Rice

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To assess the relationship among Japanese rice accessions based on SSR marker profile, a total of 64 SSR markers distributed over the 12 rice chromosomes were used (Table 2). All SSR markers were selected from a public database (http://www.gramene.org).
Genomic DNA was extracted from a young leaf from each accession. Leaf tissue was ground in 100 μl of 0.25 N NaOH with zirconium beads in 2.0-ml tubes. A volume of 400 μl of 100 mM Tris-HCl (pH 7.5) was added to each tube. The sample was then mixed and centrifuged for 10 min at 10,000 rpm. The supernatant was poured into a fresh 1.5-ml tube. PCR was performed in a 10-μl PCR mixture containing 1 μl sterile H₂O, a total of 1.5 μl forward primer (2 μM) and reverse primer (2 μM), 7.5 μl of 2× Quick Taq HS DyeMix (Toyobo Co., Ltd.), and 5 μl DNA concentrated to about 5 to 10 ng/μl. PCR amplification was performed with the following profile: 94°C for 2 min, 40 cycles of 30 s at 94°C, 30 s at 55°C, and 1 min at 68°C. To detect polymorphisms, the amplified products were separated by electrophoresis on 2% agarose gels in 1× TAE buffer at 150 V for 90 to 120 min, and the DNA fragment was detected with ethidium bromide.

Kawasaki-Tanaka A, & Fukuta Y. (2014). Genetic variation in resistance to blast disease (Pyricularia oryzae Cavara) in Japanese rice (Oryza sativa L.), as determined using a differential system. Breeding Science, 64(2), 183-192.

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Quantitative Analysis of sgRNA3 Expression

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Total RNA was extracted from cells using an RNeasy Mini Kit (Qiagen, Valencia, CA) and reverse transcribed using ReverTra Ace qPCR RT Master Mix (Toyobo, Tokyo, Japan). RT–PCR was performed to detect sgRNA3 and GAPDH mRNA by using Quick Taq HS Dye Mix (Toyobo). Quantitative RT–PCR was performed by using Fast SYBR Green Master Mix (Applied Biosystems, Foster City, CA) and a StepOne Plus Real-Time PCR System (Applied BioSystems). The expression level of sgRNA3 was calculated by the relative threshold cycle (ΔΔC_T) method using GAPDH mRNA as an internal control.
The primers for RT–PCR and qRT–PCR are listed in Table 1.

Tanaka T., Saito A., Suzuki T., Miyamoto Y., Takayama K., Okamoto T, & Moriishi K. (2022). Establishment of a stable SARS-CoV-2 replicon system for application in high-throughput screening. Antiviral Research, 199, 105268.

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Quantify CpG Methylation in EF1α Promoter

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To examine CpG methylation levels in the EF1α promoter region transduced by the pSicoR lentiviral vector, 500 ng of extracted genomic DNA was converted using the MethylEasy Xceed kit (Human Genetic Signatures). First-round PCR was performed by Quick Taq HS DyeMix (ToYoBo) using 25 ng of converted DNA in a 10 µl reaction mixture, then 0.5 µl of the first-round PCR product was used as template for nested PCR in a 25 μl reaction mixture. All primer sets and conditions are listed in Supplemental Table 6.

Yamazaki H., Shirakawa K., Matsumoto T., Hirabayashi S., Murakawa Y., Kobayashi M., Sarca A.D., Kazuma Y., Matsui H., Maruyama W., Fukuda H., Shirakawa R., Shindo K., Ri M., Iida S, & Takaori-Kondo A. (2019). Endogenous APOBEC3B Overexpression Constitutively Generates DNA Substitutions and Deletions in Myeloma Cells. Scientific Reports, 9, 7122.

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