Taq master mix

Manufactured by Vazyme

Sourced in China, United States, Germany

2 × Taq Master Mix is a ready-to-use solution that contains all the necessary components for PCR amplification, including Taq DNA polymerase, dNTPs, and buffers. It is designed to simplify the PCR setup process and improve consistency in results.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (10 mentions) T100 thermal cycler, by Bio-Rad (6 mentions) Aceq qpcr sybr green master mix, by Vazyme (5 mentions) Trizol, by Thermo Fisher Scientific (4 mentions) Hiscript 2 q rt supermix for qpcr, by Vazyme (4 mentions) Gel doc xr gel documentation system, by Bio-Rad (3 mentions) Pmd18 t, by Takara Bio (3 mentions) Hiscript 1st strand cdna synthesis kit, by Vazyme (3 mentions) Rnaiso plus, by Takara Bio (2 mentions) Maxima h minus reverse transcriptase, by Thermo Fisher Scientific (2 mentions) Primer premier 5, by Premier Biosoft (2 mentions) Penicillin and streptomycin, by Thermo Fisher Scientific (2 mentions) Az10606120, by Bio-Techne (2 mentions) Recombinant mouse scf protein, by R&D Systems (2 mentions) Salicylic acid, by Yuanye Bio-Technology (2 mentions) Nf449, by Cayman Chemical (2 mentions) 5 bdbd, by Merck Group (2 mentions) Fluo 4 am, by Solarbio (2 mentions) Trizol, by Vazyme (2 mentions) Reverse transcription kit, by Vazyme (2 mentions)

Spelling variants of this product

Taq Plus Master Mix (24 citations) EpiArt HS Taq Master Mix (5 citations) 2 × Ace Taq Master Mix (4 citations) 2 × Taq Plus Master Mix (Dye Plus) (4 citations) Taq Pro HS Universal Probe Master Mix (4 citations) Taq Master Mix (2×) (2 citations) 2 × Taq Master Mix kit (2 citations) Two × Taq Plus Master Mix II (2 citations) 3G Taq Master Mix for PAGE (2 citations) Fast-Taq Master Mix (2 citations)

154 protocols using taq master mix

Quantitative Detection of 'Candidatus Liberibacter asiaticus'

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For CLas detection, total DNA was extracted from midrib sections of sugar orange plants using Biospin Omini Plant Genomic DNA Extraction Kit (BioFlux, Hangzhou, China). The CLas detection was conducted by both PCR and quantitative PCR (qPCR). The PCR was performed using 16S rDNA primer OI1/OI2c with 2X Taq Master Mix (Vazyme, Nanjing, China) in a 20 μL reaction system. The bacterial populations (CLas cells μg^-1 of citrus DNA) were quantified with qPCR assay that was described by Du M. et al. (2021) (link). In the qPCR assay, CLasgyrA of CLas was detected, and 18S rRNA of citrus was used as the internal reference. BlastTaq™ 2X qPCR MasterMix (abm, Canada) was used for qPCR amplification. Primer pairs are listed in Table S1.
Total RNA was extracted from plant tissues and psyllids using RNAiso Plus (Takara, Japan). In a 20-μL volume, 1μg of total RNA was reverse transcribed with All-In-One 5X RT MasterMix (abm) following the manufacturer’s instructions. Quantitative RT-PCR (qRT-PCR) assays were conducted to analyze gene transcript level. Primer pairs of selected genes are listed in Table S1. The genes NbACTIN and CsGAPDH were used as endogenous controls. The qRT-PCR analyses were performed in a 10 μL reaction system. The 2^-ΔΔCt method was used for the determination of relative gene transcription (Livak and Schmittgen, 2001 (link)).

Zhang S., Wang X., He J., Zhang S., Zhao T., Fu S, & Zhou C. (2023). A Sec-dependent effector, CLIBASIA_04425, contributes to virulence in ‘Candidatus Liberibater asiaticus’. Frontiers in Plant Science, 14, 1224736.

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Quantitative RT-PCR for RUNX2 Splicing

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Total cellular RNAs were collected by total RNA miniprep kit (Axygen, USA). The cDNA was synthesized by using the Maxima H Minus Reverse Transcriptase from Thermo Fisher Scientific (USA). The following PCR reaction was performed with 2 x Taq Master Mix (Vazyme, China) using following primers: 5’-GACGAGGCAAGAGT TTCACC-3’, 5’-ATGAAATGCTTGGGAACTGC-3’, and 5’-GGTGGTAGAGTGGATGGACG-3’ for RUNX2 exon 5 and 7 alternative splicing detection, 5’-GGGAACCCAGAA GGCACAGAC-3’, 5’-GCCTGGGGTCTGTAATCTGACT C-3’ for RUNX2 exon 5, 5’-CGTCCATCCACTCTACCACC-3’, 5’-ATGAAATGCTTGGGAACTGC-3’ for RUNX2 exon 7 alternative splicing, 5’-GACAAGAAGCCCTTCACTGC-3’ and 5’-AGACTGCGCCTGGTAGTTGT-3’ for alkaline phosphatase (ALP), 5’-CGCTACCTGTATCAATGGCTG-3’ and 5’-GCCAACTCGTCACAGTCCG-3’ for osteocalcin (OC), 5’-GCTCAACCATAGAGAAAGCAAACG-3’ and 5’-CTTCGTTGCCTTTCCCAACTTC-3’ for dentin sialophosphoprotein (DSPP), 5’-GAAGGTGAAGGTCGGAGTC-3’ and 5’-GAAGATGGTGATGGGATTTC-3’ for GAPDH (Supplementary Table S1).
The accurate band intensity of RT-PCR product with local background subtraction were measured by using Quantity One software (Bio-Rad). The expression levels of a gene in each sample were normalized by GAPDH internal control. The ratio of RUNX2 exon 5 inclusion vs exclusion were calculated by dividing the band intensity of longer product by that of short product.

Shen J., She W., Zhang F., Guo J, & Jia R. (2021). YBX1 Promotes the Inclusion of RUNX2 Alternative Exon 5 in Dental Pulp Stem Cells. International Journal of Stem Cells, 15(3), 301-310.

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Quantification of Inflammatory Cytokines in BV2 Cells

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Total RNA was extracted from BV2 cells using TRIzol^® (Invitrogen; Thermo Fisher Scientific, Inc.) and reverse transcribed using the PrimeScript™ RT Master Mix (Takara Biotechnology Co., Ltd.), according to the manufacturer's protocol. Conditions for reverse transcription were 15 min at 37˚C and 5 sec at 85˚C. PCR analyses of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and β-actin mRNA levels were performed using 2X Taq Master Mix (Vazyme Biotech Co., Ltd.). The forward and reverse primer pairs used were as follows: TNF-α forward, 5'-ATCCGCGACGTGGAACTG-3' and reverse, 5'-ACCGCC TGGAGTTCTGGAA-3'; IL-1β forward, 5'-CTTCATCTT-3' and reverse, 5'-TCACACACCAGCAGGTTATCATC-3'; and β-actin forward, 5'-TGGCACCCAGCACAATGAA-3' and reverse, 5'-CTAAGTCATAGTCCGCCTAGAAGCA-3'. The PCR thermocycling conditions were as follows: 95˚C for 5 min, followed by 30 cycles of 95˚C for 30 sec, 55˚C for 30 sec and 72˚C for 1 min and 72˚C for 10 min. The products were separated on a 1.2% agarose gel containing ethidium bromide and were visualized under a gel imaging system.

Lin C., Huang Z., Zhou R., Zhou Y., Shentu Y., Yu K, & Zhang Y. (2020). Notch3 and its CADASIL mutants differentially regulate cellular phenotypes. Experimental and Therapeutic Medicine, 21(2), 117.

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Genetic Barcoding of Rehmannia Species

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Amplification primers of target sequences in Rehmannia were designed according to the appropriate region of candidate barcodes (Table S5), and target sequences were amplified by PCR according to amplification conditions in Table S6. PCR amplification volume was 25 μl, and composed of 12.5 μl reaction buffer (2xTaq Master Mix) (Vazyme, Nanjing, P. R. China), 0.5 μl each primer (10 mM), 1 μl template DNA and 10.5 μl ddH₂O. Sequencing of target sequences was performed by GENEWIZ. Inc. (Suzhou, P. R. China), DNA sequences were all submitted to GenBank (Table 1).

Duan H., Wang W., Zeng Y., Guo M, & Zhou Y. (2019). The screening and identification of DNA barcode sequences for Rehmannia. Scientific Reports, 9, 17295.

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Targeted DNA Sequencing of Key Genes

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Total DNA was extraction from cell pellets using TIANamp Genomic DNA Kit (TIANGEN). Then the genomic fragments, including PRDM15, ZBTB21, and MX1-ABCG1 were amplified using 2xTaq Master Mix (vazyme Bio-tech) with primers in table S2. PCR productions were purified and sequenced in both direction with BigDye Terminator Cycle Sequencing Kit on an ABI PRISM 3730 DNA analyser (Applied Biosystems). Sequences of each segment were proofread and assembled with DNASTAR 5.0 (DNASTAR Inc., Madison, WI, USA).

Han W., Shi D., Yang Q., Li X., Zhang J., Peng C, & Yan F. (2024). Alteration of chromosome structure impacts gene expressions implicated in pancreatic ductal adenocarcinoma cells. BMC Genomics, 25, 206.

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Amplification and Gel Electrophoresis

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Amplification was conducted using 2X Taq Master Mix (Vazyme, Nanjing, China). The resulting products underwent agarose gel electrophoresis, and outcomes were observed and captured using a gel imaging system.

Liang W., Zhou Z., Gao Q., Zhu Z., Zhu J., Lin J., Wen Y., Qian F., Wang L., Zhai Y., Lv J., Zhang H., Zhong F, & Du H. (2024). Tumor‐derived Prevotella intermedia aggravates gastric cancer by enhancing Perilipin 3 expression. Cancer Science, 115(4), 1141-1153.

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Reverse Transcription and PCR for Splicing Analysis

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The cDNA was reverse-transcribed from the total RNA using Maxima H Minus Reverse Transcriptase (Thermo Fisher Scientific, USA). PCR amplification was performed with 2 x Taq Master Mix (Vazyme, NJ, USA) using the listed primers: 5′-ACGCCATCTTTCAGAACTGTGCT-3′ and 5′-CATATTCTGCGGGGTGATCTC-3′ for hnRNP L exon 7 inclusion detection, 5′-ATCAGGATACTTGGGACTACACAAAC-3′ and 5′-CATCATGGTAATGGCTGTGGTAC-3′ for hnRNP L exon 7 exclusion and inclusion detection, 5′-GGAGTCCTCCACCTCGTCGCA-3′ and 5′-ACGAGACCTAGAGAAGGATCGGGAC-3′ for SRSF3 exon 4 splicing detection, and 5′-GAAGGTGAAGGTCGGAGTC-3′ and 5′-GAAGATGGTGATGGGATTTC-3′ for GAPDH.

Xu L., Shen J., Jia J, & Jia R. (2019). Inclusion of hnRNP L Alternative Exon 7 Is Associated with Good Prognosis and Inhibited by Oncogene SRSF3 in Head and Neck Squamous Cell Carcinoma. BioMed Research International, 2019, 9612425.

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Mosquito Genomic DNA Extraction and PCR

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Genomic DNA was extracted by homogenizing individual female mosquitoes in 200 μL STE buffer with a sterile pestle, followed by a 30 min incubation at 65°C with 2 μL of Proteinase K (20 mg/mL) and a final incubation for 10 min at 95°C. The extracted DNA was diluted 1/5 in water and samples were centrifuged at 2,000 g for 2 min before PCR. For each PCR reaction, 2 μL of DNA template was amplified using the 2x Taq master mix (Vazyme) in a 25 μL reaction: 12.5 μL of master mix, 12.5 μL of water, 1 μL of each 10 μM primer (S5 Table) and the following PCR cycles: 95°C for 3 min, 35 cycles of 15s denaturation at 95°C, 15s for primer annealing (see temperatures in S5 Table), 1 min extension at 72°C and a 5 min final extension step at 72°C. Wolbachia density relative to host DNA and plasmid copy number were measured by qPCR using the QuantiNova SYBR Green PCR kit (Qiagen) in 10 μL reactions: 5 μL of master mix, 2 μL of water, 0.5 μL of each 5 μM primer (S5 Table) and the following cycles: 95°C for 15 min, 40× cycles of 95°C for 15 s and 60°C for 20 s, followed by a melt-curve analysis.

Martinez J., Ant T.H., Murdochy S.M., Tong L., da Silva Filipe A, & Sinkins S.P. (2022). Genome sequencing and comparative analysis of Wolbachia strain wAlbA reveals Wolbachia-associated plasmids are common. PLoS Genetics, 18(9), e1010406.

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PCR Detection of Antibiotic Resistance Genes

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Antibiotic resistance genes (mecA and mecC) were amplified by using a basic Thermal Cycler PCR (BIO-RAD T100^TM Thermal Cycler, Massachusetts, USA) in which specific primers were used for both mecA and mecC genes (Table 1). A total volume of the PCR reaction (15 µL) contained 2 µL (10 ng/µL) of DNA, 7.5 µL of 2X Taq master mix (Vazyme Biotech Co., Nanjing, China), 1 µL (10 µM) of forward primer, 1 µL (10 µM) of reverse primer, and 3.5 µL of deionized water. PCR for both genes (mecA and mecC genes) was performed by applying the following conditions: first denaturation at 95 °C for 5 min followed by 35 cycles of 95 °C for 1 min, 55 °C for 30 s, 72 °C for 1 min, and 72 °C for 5 min [21 (link),22 (link)]. Molecular detection of mecA and mecC was performed by loading 6–7 µL of 100 bp DNA ladder into the first well and the remaining wells were loaded by PCR products using 1.5% agarose gel containing 0.5 mg/mL of ethidium bromide in Tris-borate-EDTA (10 mM+1mM EDTA; pH 8.0) buffer used by applying 120 V for an hour. DNA bands were observed under ultraviolet light (UV-light) using the Gel Doc system (BIO-RAD Gel Doc^TM XR + with Image Lab^tm Software, BioRad, Hercules, CA, USA).

Idrees M.M., Saeed K., Shahid M.A., Akhtar M., Qammar K., Hassan J., Khaliq T, & Saeed A. (2023). Prevalence of mecA- and mecC-Associated Methicillin-Resistant Staphylococcus aureus in Clinical Specimens, Punjab, Pakistan. Biomedicines, 11(3), 878.

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Chicken IGF-I Gene Amplification

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Based on the chicken IGF-I gene sequence (GenBank accession No. NC_006088.4, Gene ID 418090), the Primer Premier 5 ® software (Premier Biosoft, Palo Alto, CA, USA) was used to design three pairs of primers to amplify parts of the exon regions of the IGF-I gene. For expression profile analysis, two pairs of primers were designed to perform qPCR (Table 1). All primers were synthesized by Sangon Biotech (Shanghai) Co., Ltd. (Shanghai, China). The PCR was performed in a 20-µL reaction volume comprising 1.5 µL chicken genomic DNA (50 ng/µL), 10 µL 2X Taq Master Mix (Vazyme Biotech Co., Ltd, Nanjing, China), 0.4 µL forward primer (10 µM); 0.4 µL reverse primer (10 µM), and 7.7 µL sterilized distilled water. The following amplification conditions were used: initial denaturation at 94°C for 7 min; followed by 35 cycles of denaturation at 94°C for 30 s, annealing at 59°C for 30 s, and extension at 72°C for 30 s; and final extension at 72°C for 10 min. The PCR products were verified by 10% non-denaturing polyacrylamide gel electrophoresis.

Abdalhag M.A., Li T., Duan L., Zhang T., Zhang G., Wang J, & Wang Y. (2016). Association analysis of IGF-I gene expression with growth and reproductive traits in Jinghai yellow chickens. Genetics and molecular research : GMR, 15(4).

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