Taq pcr starmix with loading dye

Manufactured by GenStar

Sourced in China

2× Taq PCR StarMix with Loading Dye is a ready-to-use PCR master mix that contains Taq DNA polymerase, dNTPs, and necessary buffers. The mix also includes a loading dye for direct gel loading after the PCR reaction.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (4 mentions) M mlv reverse transcriptase, by Promega (3 mentions) Abi 3730 dna analyzer, by Thermo Fisher Scientific (3 mentions) Rnase free dnase 1, by Takara Bio (2 mentions) Ultrasybr mixture, by CWBIO (2 mentions) T100 thermal cycler, by Bio-Rad (2 mentions) Taq master mix, by Vazyme (1 mentions) Lightcycler480 2 rt pcr detection system, by Roche (1 mentions) Lightcycler 480 2, by Roche (1 mentions) Dnase 1, by Takara Bio (1 mentions) Qtower 2.2 real time pcr system, by Analytik Jena (1 mentions) Realstar green fast mixture with rox 2, by GenStar (1 mentions) Star script 2 first strand cdna synthesis kit with gdna remover, by GenStar (1 mentions) Plant rna kit, by Omega Bio-Tek (1 mentions) Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (1 mentions) T gradient thermoblock, by Analytik Jena (1 mentions) Gelview, by BioTeke (1 mentions)

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2×Taq PCR StarMix (31 citations)

12 protocols using taq pcr starmix with loading dye

Multiplex PCR for Feline Viral Pathogens

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To establish the mPCR methods, two mixtures of plasmid standards were used to verify the optimum annealing temperature and the ratio of each primer. The reaction system volume for PAIT was 20 µL, including 10 µL of 2× Taq Master Mix (Vazyme, Nanjing, China), 1 µL of mixed forward primers (FCoV-F(0.26 µL), FeAstV-F(0.16 µL), FPV-F(0.16 µL) and FeKoV-F(0.42 µL)), 1µL of mixed reverse primers (FCoV-R(0.26 µL), FeAstV-R (0.16 µL), FPV-R (0.16 µL) and FeKoV-R (0.42 µL)) and 1 µL of 10¹⁰ copies/µL of the mixed template (18-T-FCoV, 18-T-FeAstV, 18-T-FPV and 18-T-FeKoV). The reaction system volume for PAFR was 30 µL, consisting of 15 µL of Taq PCR StarMix with Loading Dye (GenStar, Beijing, China), 1 µL of mixed forward primers (FCV-F (0.13 µL), FHV-1-F (0.10 µL), FeLV-F (0.24 µL), C.felis-F (0.19 µL) and IAV-F (0.34 µL)), 1 µL of mixed reverse primers (FCV-R (0.13 µL), FHV-1-R (0.10 µL), FeLV-R (0.24 µL), C.felis-R (0.19 µL) and IAV-R (0.34 µL)), 1 µL of 10¹⁰ copies of the mixed template (18-T-FCV, 18-T-FHV-1, 18-T-FeLV, 18-T-C.felis and 18-T-IAV) and 12 µL ddH2O. Both mPCR procedures were as follows: 98 °C for 3 min; 36 cycles of 98 °C for 20 s, gradient annealing temperature of 50~60 °C for 1 min and 72 °C for 1 min; and 72 °C for 5 min.
The products of mPCR (15 μL) were detected with 2% agarose gel electrophoresis.

Xiao X., Hao X., Chen B., Zhou P, & Li S. (2022). Two Multiplex PCR Methods for Detecting Several Pathogens Associated with Feline Respiratory and Intestinal Tracts. Veterinary Sciences, 10(1), 14.

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Foxtail Millet and Arabidopsis RNA Extraction and Expression Analysis

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Total RNA from foxtail millet and Arabidopsis was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA). After digestion with RNase-free DNase I (Takara, Dalian, China), 2 µg of total RNA was converted into cDNA by M-MLV Reverse Transcriptase (Promega, Madison, WI, USA).
The reverse transcription polymerase chain reaction (RT-PCR) was performed using 2× Taq PCR StarMix with Loading Dye (GenStar, Beijing, China). PCR reactions were 95°C for 3 min, followed by 95°C for 30 sec, 60°C for 30 sec, 72°C for 30 sec for 25 cycles and 72°C for 5 min. Primers are listed in Table S1.
A quantitative real-time PCR (qRT-PCR) assay was performed using a LightCycler 480 II RT-PCR detection system (Roche, USA) with the UltraSYBR reagent mixture (CWBIO, Beijing, China). The PCR conditions were 95°C for 10 min, followed by 40 cycles of 95°C for 15 sec, 60°C for 1 min. The relative expression levels of mRNA were calculated using the ΔΔC_T method.

Yue J., Li C., Liu Y, & Yu J. (2014). A Remorin Gene SiREM6, the Target Gene of SiARDP, from Foxtail Millet (Setaria italica) Promotes High Salt Tolerance in Transgenic Arabidopsis. PLoS ONE, 9(6), e100772.

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Quantitative RT-PCR Gene Expression Analysis

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Total RNA was isolated using TRIzol reagent (Invitrogen, USA). After digestion with RNase-free DNase I (Takara, Japan), ~2 μg of total RNA was used for reverse transcription by M-MLV Reverse Transcriptase (Promega, USA).
Reverse transcription polymerase chain reaction (RT-PCR) was performed using 2 × Taq PCR StarMix with Loading Dye (GenStar, China). PCR conditions were 95°C for 3min, followed by 24 cycles of 95°C for 30 s, 60°C for 30 s, 72°C for 30 s, and 72°C for 5min. Quantitative RT-PCR (qRT-PCR) assays were performed with a LightCycler 480 II real-time PCR detection system (Roche, USA) using the UltraSYBR Mixture (CWBIO, China). The PCR conditions were 95°C for 10min, followed by 40 cycles of 95°C for 15 s, and 57°C or 60°C for 1min. The ΔΔC_T method was used to calculate the expression levels of relevant genes.

Li C., Yue J., Wu X., Xu C, & Yu J. (2014). An ABA-responsive DRE-binding protein gene from Setaria italica, SiARDP, the target gene of SiAREB, plays a critical role under drought stress. Journal of Experimental Botany, 65(18), 5415-5427.

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Characterization of maize mutant mesocotyl

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The zmnac17-1 and zmnac17-2 mutants were collected from the maize EMS mutant library [41 (link)]. The B73 wild type from the same library was used as a control. All materials were propagated in Jiaozhou, Shandong (36°27′ N, 120°03′ E) in summer and in Ledong, Hainan (18°45′ N, 109°10′ E) in winter. Genotyping was evaluated using Sanger sequencing. DNA from leaves was extracted using the CTAB method. Primers were designed using the NCBI website (Supplementary Table S1). PCR was performed using 2× Taq PCR StarMix with Loading Dye (A012, GenStar). Sanger sequencing was performed by Sangon Biotech Co., Ltd (Shanghai, China).
zmnac17-1, zmnac17-2, and B73 were used for mesocotyl elongation analysis. zmnac17-1 and B73 were used for physiological and biochemical analyses, hormone content determination, and RNA sequencing analysis. Seeds were sown in a 54 × 28 × 9 cm high-footed seedling tray in the dark. Each hole was filled with vermiculite and then fully watered, and the seedlings were grown in a dark incubator (25 °C). The mesocotyl lengths of the 7-day-old seedlings were measured according to previously published methods [5 (link)]. At least 15 individual seedlings for each genotype were analyzed.

Yang R., Li K., Wang M., Sun M., Li Q., Chen L., Xiao F., Zhang Z., Zhang H., Jiao F, & Chen J. (2024). ZmNAC17 Regulates Mesocotyl Elongation by Mediating Auxin and ROS Biosynthetic Pathways in Maize. International Journal of Molecular Sciences, 25(9), 4585.

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ITS Sequencing of Edgeworthia Species

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Polymerase chain reaction (PCR) amplification was conducted on a final reaction volume, with a 20 μL volume reaction consisting of 10 μL of 2× Taq PCR StarMix with loading dye (Genstar Biosolutions, China), 1 μL of each primer, 6 μL of distilled water, and 2 μL 5 ng genomic DNA as a template. The ITS universal primer set: ITS1, 5′-TCC GTA GGT GAA CCT GCG G-3′ (forward) and ITS4, 5′-TCC TCC GCT TAT TGA TAT GC-3′ (reverse), was used to obtain the ITS region (White et al., 1990 (link)). PCR amplification was programmed with thermal settings of an initial denaturation at 94°C for 5 min; denaturation at 94°C for 60 s, annealing at 55°C for 60 s, extension at 72°C for 60 s; and a final extension at 72°C for 7 min. Upon verification via electrophoresis on a 1.0% agarose gel and documented under the UV machine, the PCR products were sent for direct Sanger sequencing at both ends using an ABI 3730 DNA Analyzer (Applied Biosystems, United States). Results acquired from the Sanger sequencing were aligned and manually edited to obtain the clean sequences of the three species of Edgeworthia. The ITS sequences for E. albiflora (MW255615), E. chrysantha (MW255616), and E. gardneri (MW255617) were deposited in the NCBI GenBank database for future reference.

Qian S., Zhang Y, & Lee S.Y. (2021). Comparative Analysis of Complete Chloroplast Genome Sequences in Edgeworthia (Thymelaeaceae) and New Insights Into Phylogenetic Relationships. Frontiers in Genetics, 12, 643552.

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Gene Expression Analysis in Foxtail Millet and Arabidopsis

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Total RNA was extracted from foxtail millet and Arabidopsis using the TRIzol reagent (Invitrogen, USA). After digestion with DNaseI (Takara, Japan), 3–5 μg of total RNA was prepared, and cDNA was synthesized via reverse transcription using M-MLV Reverse Transcriptase (Promega, USA). Semi-quantitative RT-PCR was performed using 2 × Taq PCR StarMix with Loading Dye (GenStar, China). The PCR conditions were 95°C for 5 min, followed by 25 cycles of 95°C for 30 s, 60°C for 30 s, and 72°C for 30 s, with a final step at 72°C for 10 min. Quantitative RT-PCR (qRT-PCR) was performed using 2 × Ultra SYBR Mixture (CWBIO, China) on a qTower 2.2 Real-Time PCR System (AnalytikJena, Germany). The PCR conditions were 95°C for 10 min, followed by 40 cycles of 95°C for 15 s, and 60°C for 1 min. Relative gene expression levels were calculated using the 2^−ΔΔCT method (Livak and Schmittgen, 2001 (link)).

Li J., Dong Y., Li C., Pan Y, & Yu J. (2017). SiASR4, the Target Gene of SiARDP from Setaria italica, Improves Abiotic Stress Adaption in Plants. Frontiers in Plant Science, 7, 2053.

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Sanger Sequencing of nrITS Region

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Three to four biological replicates from each species were selected for Sanger sequencing to obtain their nrITS sequences. PCR amplification of the nrITS region was carried out using primers ITS-p5: 5′-CCT TAT CAY TTA GAG GAA GGA G-3’ (Cheng et al., 2016 (link)) and ITS-S3R: 5′-GAC CGT TCT CCA GAC TAC AAT-3’ (Chiou et al., 2007 (link)). PCR was conducted in a final reaction volume of 20 μL, containing 10 μL of 2 × Taq PCR Starmix with loading dye (Genstar Biosolutions, China), 0.4 μM of each primer, and 20 ng of genomic DNA as template. PCR amplification was conducted in a T100™ Thermal Cycler (Bio-Rad, USA), with thermal settings programmed as follows: initial denaturation at 94 °C for 5 min; 40 cycles of 94 °C for 30 s, 55 °C for 30 s, and 72 °C for 30 s; and a final extension at 72 °C for 7 min. PCR products were sent for direct sequencing for both ends on an ABI 3730 DNA Analyzer (Applied Biosystems, USA).

Lee S.Y., Xu K.W., Huang C.Y., Lee J.H., Liao W.B., Zhang Y.H, & Fan Q. (2021). Molecular phylogenetic analyses based on the complete plastid genomes and nuclear sequences reveal Daphne (Thymelaeaceae) to be non-monophyletic as current circumscription. Plant Diversity, 44(3), 279-289.

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Total RNA Extraction and RT-qPCR Analysis

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Total RNA was extracted using the Plant RNA kit (Omega Bio-Tek, Norcross, GA, USA). The optical density of the RNA extracts was determined using an ultraviolet spectrophotometer, and total RNA purity was estimated by calculating OD₂₆₀/OD₂₈₀, which remained within 1.8 and 2.2. The concentration of total RNA was calculated according to the dilution ratio and the value of OD₂₆₀.
cDNAs were synthesized using a StarScript II First-Strand cDNA Synthesis Kit with gDNA Remover (GenStar, Beijing, China). PCR amplifications were carried out using a 2× Taq PCR StarMix with Loading Dye (GenStar) and run according to the program as described (a. 2 min at 94 °C; b. 30 s at 94 °C; c. 30 s at 55 °C; d. 1 min at 72 °C; e. Repeat steps b–d for optimized cycles ranging from 12 to 28; f. 8 min at 72 °C; g. Hold at 4 °C). The gene product was electrophorized in 1% agarose gel. RT-qPCR was carried out using a 2× RealStar Green Fast Mixture with ROX II (GenStar) and run according to the program as described (a. 2 min at 95 °C; b. 15 s at 95 °C; c. 25 s at 55 °C; d. 30 s at 72 °C; e. Repeat steps b–d for 40 cycles).
The specific primers to selected genes were designed using Primer 5.0 software, and the sequences of the primers used are shown in Table 1 and Table 2.

Tan Q., Zhu J., Ju Y., Chi X., Cao T., Zheng L, & Chen Q. (2023). Antiviral Activity of Ailanthone from Ailanthus altissima on the Rice Stripe Virus. Viruses, 16(1), 73.

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PCR Amplification of Nuclear Ribosomal ITS

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Polymerase chain reaction (PCR) amplification was carried out in a 20 µL reaction volume using the ITS universal primer set: 5F: 5ʹ-GGAAGTAAAAGTCGTAA-CAAGG-3ʹ (forward) and 4R: 5ʹ-TCCTCCGCTTATTGATATGC-3ʹ (reverse). The PCRs for the nuclear ribosomal DNA ITS region contained 10 µL of 2× Taq PCR Starmix with loading dye (Genstar Biosolutions, China), 0.4 µM of each primer and 20 ng of genomic DNA as a template. PCR amplifications were conducted on a T100 Thermal Cycler (Bio-Rad, USA), with initial denaturation at 93 °C for 5 min; 40 cycles of denaturation at 93 °C for 30 s, annealing at 60 °C for 30 s, and extension at 72 °C for 30 s; and a final extension at 72 °C for 5 min. PCR products were sent for direct Sanger sequencing at both ends using an ABI 3730 DNA Analyzer (Applied Biosystems, USA).

He L., Zhang Y, & Lee S.Y. (2021). Complete plastomes of six species of Wikstroemia (Thymelaeaceae) reveal paraphyly with the monotypic genus Stellera. Scientific Reports, 11, 13608.

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Generating Homozygous Transgene-Free Mutants

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To obtain homozygous transgene-free mutants, DNA and proteins of T 1 seedlings were extracted for testing. At least ve T1 plants from each independent T 0 plant were selected for further analysis of sitespeci c mutations. To analyze the knock-out mutation genotypes of the T 1 plants, all transgenic lines were detected by PCR ampli cation using a pair of primers (JD-F3 and JD-R508) surrounding the target region of OsCSN1. The PCR products were generated using a 2×Taq PCR StarMix with Loading Dye (GenStar, China), and sequenced to determine the mutation sites of OsCSN1. Two transgene-free mutant lines oscsn1-580 and oscsn1-191) were successfully identi ed (Fig. 2). The T 2 plants and advancedgeneration seeds were used for the following experiments.

Han S., Liu Y., Xu M., Gao X., Zhang C., Chen S., Wu M., & Guo L. (2021). Generation and Characterization of oscsn1 Mutants Reveal That OsCSN1 Regulate ABI5 Degradation In Seedling Growth.

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