Prime taq dna polymerase

Manufactured by GeNet Bio

Prime Taq DNA polymerase is a thermostable DNA polymerase enzyme used for DNA amplification in polymerase chain reaction (PCR) applications. It catalyzes the synthesis of DNA from nucleotide precursors.

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Lab products found in correlation

Prime taq buffer, by GeNet Bio (3 mentions) Dntps mixture, by GeNet Bio (3 mentions) Ab 2720 thermocycler, by Thermo Fisher Scientific (3 mentions) Vg sys horizontal electrophoresis system, by Harvard Bioscience (3 mentions) Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions) Mluci, by New England Biolabs (1 mentions) Abi 3730xl dna analyzer, by Thermo Fisher Scientific (1 mentions) Piko thermal cycler, by Thermo Fisher Scientific (1 mentions) Primeprep pcr purification kit, by GeNet Bio (1 mentions) Restriction enzyme, by New England Biolabs (1 mentions) Pikoreal real time pcr system, by Thermo Fisher Scientific (1 mentions) Sybr green realtime pcr master mix, by Toyobo (1 mentions) Quantitect reverse transcription kit, by Qiagen (1 mentions) Abi 3130 genetic analyzer, by Thermo Fisher Scientific (1 mentions) Qiaquick pcr purification kit, by Qiagen (1 mentions) Dna thermal cycler, by Eppendorf (1 mentions)

Spelling variants of this product

Taq DNA polymerase (16 citations) Taq polymerase (13 citations) Prime Taq polymerase (5 citations) HS Prime Taq DNA Polymerase (4 citations) Prime TaqTM DNA Polymerase (3 citations) Prime Taq DNA Polymerase Kit (2 citations)

9 protocols using prime taq dna polymerase

Genetic Profiling of Leptin and BMP4

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Genomic DNA was extracted from the peripheral blood leucocytes using magnetic bead technology (chemagic DNA Blood Kit special, Chemagen, Germany) on automated high throughput nucleic acid isolation platform (chemagic Magnetic Separation Module I, Chemagen, Germany).
The polymorphic regions of leptin and BMP4 genes were amplified by polymerase chain reaction (PCR) with primer sets previously designed by Mórocz et al. [20 (link)]. The PCR was carried out in a reaction mix of 20 μL containing 100 ng DNA and 10x Prime Taq buffer (Genet Bio, Korea), 10 mM dNTPs mixture (Genet Bio, Korea), 20 pmol forward and reverse primers (AlphaDNA, Canada), and 0.1 U Prime Taq DNA polymerase (Genet Bio, Korea). PCR amplifications were performed in an AB 2720 Thermocycler (Life Technologies, USA) with an initial denaturation at 94°C for five minutes and a final extension of seven minutes at 72°C. The following thermal cycle was repeated 30 times: denaturation at 94°C for 30 seconds, annealing at 55°C for leptin and 59°C for BMP4 for 30 seconds, and extension at 72°C for 30 seconds.
The PCR products were cleaved with the restriction enzymes: HhaI (NEB, USA) for leptin and MluCI (NEB, USA) for BMP4, according to the manufacturer's instructions, and the obtained fragments were separated on agarose 3% gel in VG-SYS Horizontal Electrophoresis System (Biochrom, UK).

Nikolova S., Yablanski V., Vlaev E., Getova G., Atanasov V., Stokov L., Savov A.S, & Kremensky I.M. (2015). In Search of Biomarkers for Idiopathic Scoliosis: Leptin and BMP4 Functional Polymorphisms. Journal of Biomarkers, 2015, 425310.

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cDNA Synthesis and GAPDH Amplification

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Complementary DNA (cDNA) synthesis was performed in a total volume of 20 μl, containing 200 U of M-MuLV Reverse Transcriptase, 4 μl of 5 × Reaction Buffer, 20 U of Ribolock™ RNase inhibitor, 2 μl of Deoxyribonucleotide triphosphate (final 1 mM), 1 μl of oligo (dt) 18 primer (0.5 μg), and 1 μg of total RNA. The prepared template with the control Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) RNA and GAPDH control primers (1.3 kb) were used as a positive control, and no negative template control was prepared with total reagent for the reverse transcription reaction, except for the RNA template. cDNA was synthesized according to the manufacturer recommendations (RevertAid™ First Strand cDNA Synthesis Kit, Fermentas). Each cDNA was amplified by control polymerase chain reaction (PCR) reaction using primers for the GAPDH according to the manufacturer protocol (Prime Taq DNA polymerase, Genet Bio, South Korea). Five microliter of PCR product was loaded on 1% agarose gel and a distinct 496 bp was observed after Ethidium Bromide staining.

Azimian H., Bahreyni-Toossi M.T., Rezaei A.R., Rafatpanah H., Hamzehloei T, & Fardid R. (2015). Up-regulation of Bcl-2 expression in cultured human lymphocytes after exposure to low doses of gamma radiation. Journal of Medical Physics / Association of Medical Physicists of India, 40(1), 38-44.

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Mitochondrial Gene Amplification in Pituophis kaibarae

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We used PCR (polymerase chain reaction) to amplify a partial fragment of the mitochondrial control region (CR), Cyt b (cytochrome b), and COI (cytochrome oxidase subunit I) from a total of 128 individuals in eight P. kaibarae populations (Table S1) using previously published primers (see Table S2 for detailed information), CR (L-Thr and H-12S; Takahashi and Goto 2001 (link)), Cyt b (PunCytBF and PunCytBR; Miya et al. 2001 (link)), and COI (HCO and LCO; Folmer et al. 1994 (link)). PCR was performed using a Piko Thermal Cycler (Finnzymes, Espoo, Finland) with a 25-μL reaction mixture containing 1 μL genomic DNA, 1X Taq buffer, 0.2 mmol/L dNTP, 0.25 μmol/L of each primer, and 2.5 unit of Prime Taq DNA polymerase (GenetBio, Daejeon, South Korea). Thermal cycling consisted of a preliminary denaturation step at 94°C for 10 min followed by 35 cycles of 94°C for 30 sec, 54–58°C for 30 sec, and 72°C for 30 sec and final extension at 72°C for 10 min. PCR products were purified using a Primeprep PCR Purification Kit (GenetBio) for direct sequencing. The amplified products were sequenced by Genotech (Deajeon, South Korea) on an ABI 3730XL DNA Analyzer (Applied Biosystems, Foster City, CA).

Bae H.G, & Suk H.Y. (2015). Population genetic structure and colonization history of short ninespine sticklebacks (Pungitius kaibarae). Ecology and Evolution, 5(15), 3075-3089.

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Genotyping IL-6 Gene Polymorphism

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Genomic DNA was extracted from the peripheral blood leucocytes using magnetic bead technology (chemagic DNA Blood Kit special, Chemagen) on automated high throughput nucleic acid isolation platform (chemagic Magnetic Separation Module I, Chemagen).
The polymorphic region of the IL-6 gene was amplified by polymerase chain reaction (PCR). The primer set [5 (link)] is listed in Table 1.
The PCR was carried out in a reaction mix of 20 μL containing 100 ng DNA and 10x Prime Taq buffer (Genet Bio), 10 mM dNTPs Mixture (Genet Bio), 20 pmol Forward and Reverse primers (AlphaDNA), and 0.1 U Prime Taq DNA Polymerase (Genet Bio). PCR amplification was performed in an AB 2720 Thermocycler (Life Technologies) with an initial denaturation at 94°C for five minutes and a final extension of seven minutes at 72°C. The following thermal cycle was repeated 30 times: denaturation at 94°C for 30 seconds, annealing for 30 seconds at temperature presented in Table 2, and extension at 72°C for 30 seconds.
The PCR product was cleaved with SfaNI restriction endonuclease (New England Biolabs), according to the manufacturer's instructions, and the restriction fragments were separated on agarose 3% gel in VG-SYS Horizontal Electrophoresis System (Biochrom). The lengths of the fragments representing the genotypes of IL-6 are presented in Table 2.

Nikolova S., Dikova M., Dikov D., Djerov A., Dzhebir G., Atanasov V., Savov A, & Kremensky I. (2015). Role of the IL-6 Gene in the Etiopathogenesis of Idiopathic Scoliosis. Analytical Cellular Pathology (Amsterdam), 2015, 621893.

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Genotyping of Polymorphic DNA Regions

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Genomic DNA was extracted from the peripheral blood leucocytes using magnetic bead technology (chemagic DNA Blood Kit special, Chemagen) on automated high throughput nucleic acid isolation platform (chemagic Magnetic Separation Module I, Chemagen).
The polymorphic regions of the genes were amplified by polymerase chain reaction (PCR). The primer sets are listed in Table 1.
The PCR was carried out in a reaction mix of 20 µL containing 100-ng DNA and 10x Prime Taq buffer (Genet Bio), 10 mM dNTPs Mixture (Genet Bio), 20 pmol Forward and Reverse primers (Alpha DNA), and 0.1 U Prime Taq DNA Polymerase (Genet Bio). PCR amplification was performed in an AB 2720 Thermocycler (Life Technologies) with an initial denaturation at 94°C for five minutes and a final extension of seven minutes at 72°C. The following thermal cycle was repeated 30 times: denaturation at 94°C for 30 seconds, annealing for 30 seconds at temperature presented in Table 2, and extension at 72°C for 30 seconds.
The PCR products were cleaved with the appropriate restriction enzymes (New England Biolabs), according to the manufacturer's instructions, and the restriction fragments were separated on agarose 3% gel in VG-SYS Horizontal Electrophoresis System (Biochrom). The restriction enzymes and the lengths of the fragments representing the genotypes are presented in Table 2.

Nikolova S., Yablanski V., Vlaev E., Stokov L., Savov A.S, & Kremensky I.M. (2015). Association Study between Idiopathic Scoliosis and Polymorphic Variants of VDR, IGF-1, and AMPD1 Genes. Genetics Research International, 2015, 852196.

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Isolation of Temperature-Sensitive OLE1 Mutants

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The OLE1 sequence was amplified by error-prone PCR using pRS414-OLE1 as the template in a 50 µl reaction containing 1 µM each of T3 and T7 primers, 1× Prime Taq reaction buffer (GeNet Bio), 0.2 mM dNTP, 0.1 mM MnCl₂, and 1 unit of Prime Taq DNA Polymerase (GeNet Bio). The PCR product was purified and transformed into ole1Δ::Kan strain together with pRS416-OLE1 cut with EcoRI to remove the coding sequence. The transformants carrying gap-repaired pRS416-OLE1 plasmids were selected on CSM-URA plates containing 2% glucose at room temperature (RT, ∼25–27°C). The plates of ∼5,000 colonies in total were replicated and placed at 37°C. The colonies that grew at RT, but not at 37°C, were individually streaked out to confirm their temperature sensitivity and their growth on selection plates containing OA. The gap-repaired pRS416 plasmids carrying ole1 mutants were isolated from cells, tested again for the temperature sensitivity, and sequenced. The ole1^ts sequence was isolated by cutting the plasmid with NotI and XhoI and used to transform ole1Δ::Kan cells. Transformants were selected for growth on YPD plates without OA and kanamycin at RT.

Huang L.J, & Chen R.H. (2022). Lipid saturation induces degradation of squalene epoxidase for sterol homeostasis and cell survival. Life Science Alliance, 6(1), e202201612.

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Semiquantitative and Quantitative RT-PCR Analysis of Citrus OMT Genes

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The first-strand cDNA was synthesized from 1 μg of total RNA in 20 μL of a reaction mixture using a QuantiTect Reverse Transcription Kit (Qiagen) according to the manufacturer's instructions. Semiquantitative RT-PCR for CreOMT1, CreOMT2, and CreOMT4 was performed with 1 μL of the first-strand cDNA as a template in a total volume of 20 μL using PrimeTaq DNA Polymerase (GENETBIO Inc., Daejeon, Korea) with gene-specific primer sets (Table 2). CuActin (C. unshiu actin) was used as an internal control. The RT-PCR products were separated by 2% agarose gel. Further expression analysis of CreOMT1 and CreOMT4 was carried out with qRT-PCR. One microliter of the first-strand cDNA was used as a template in a total volume of 12.5 μL using SYBR ® Green Real-Time PCR Master Mix (TOYOBO). The qRT-PCR was performed using a Thermo Scientific TM PikoReal TM Real-Time PCR System (Thermo Fisher Scientific Inc., Waltham, MA, USA) as follows: 95°C for 1 min, 40 cycles of 95°C for 15 s, and 60°C for 1 min for CreOMT1, CreOMT4, and CuActin. CuActin was used as a reference gene for normalization of the transcript levels of CreOMT1 and CreOMT4, as described in Kotoda et al. (2016) . Three technical replicates were performed for each reaction. The primer sets used in the qRT-PCR are listed in Table 2.

Zohra F.T., Takematsu S., Itami Y., & Kotoda N. (2020). Accumulation of Polymethoxyflavones and <i>O</i>-methyltransferase Gene Expression in Various Citrus Cultivars. The Horticulture Journal, 89(3), 225-236.

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Reverse Transcription and cDNA Synthesis

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All reactions were carried out in add up to volume of 20 μL. Each reaction mix contained 200 U of M-MuLV Reverse Transcriptase, 4 μl of 5 × Reaction Buffer, 20 U of Ribolock™ RNase inhibitor, 2 μl of Deoxyribonucleotide triphosphate (final 1 mM), 1 μl of oligo (dt) 18 primer (0.5 μg) and 1 μg of total RNA. cDNA was synthesized according to the manufacturer recommendations (RevertAid™First Strand cDNA Synthesis Kit, Fermentas). Each cDNA was confirmed by control polymerase chain reaction (PCR) reaction using primers for the glyceraldehyde 3-phosphate dehydrogenase according to the manufacturer protocol (Prime Taq DNA polymerase, Genet Bio, South Korea). The PCR product (5 μl) was loaded on a 1% agarose gel, stained with ethidium bromide, and observed with an ultraviolet transilluminator.

Toossi M.T.B., Azimian H., Soleymanifard S., Vosoughi H., Dolat E., Rezaei A.R, & Khademi S. (2020). Regulation of XPA could play a role in inhibition of radiation-induced bystander effects in QU-DB cells at high doses. Journal of cancer research and therapeutics, 16(Supplement).

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Rifampin Resistance Cluster Amplification

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The oligonucleotide primers 5ꞌ-TCT TTA TGG GTG ATT TCC CA-3ꞌ and 5ꞌ-ACC GTC GTT TAC GTT CTG TA-3ꞌ (Didier et al., 2011) were utilized for the amplification of rifampin resistance clusters I and II encompassing the rpoB gene sequence. Polymerase chain reaction (PCR) was carried out in a 50-μL volume containing 6.25 μL 0.35 µM of each primer, 6.25 μL 0.25 mM of each dNTP (dATP, dCTP, dGTP, and dTTP), 2.5 μL 100 ng DNA tem-plate, 1.5 μL 1.5 mM MgCl 2 , 0.55 μL 1U Prime Taq DNA polymerase (Genetbio, Daejeon, Korea), 5 μL 1X Taq buffer, and 21 μL MilliQ water. The PCRs were performed in a DNA thermal cycler (Eppendorf; Hamburg, Germany). The PCR cycling programs consisted of an initial denaturation (4 min at 94°C) followed by 40 cycles of denaturation (1 min at 94°C), annealing (1 min at 55°C), and extension (1 min at 72°C), with a final extension (7 min at 72°C). The amplicon of the partial cluster I gene was purified using a QIAquick PCR purification kit (Qiagen, Madrid, Spain). The sequences of the purified amplicons were obtained by Sanger dideoxy sequencing, carried out by Ocimum Biosolution (HITEC City, India) using an ABI 3130 genetic analyzer (Applied Biosystems, Foster City, CA, USA). The rpoB sequence was submitted to the National Center for Biotechnology Information (NCBI) and has been assigned the accession No. KJ559410.

Murugan K., Kavitha K, & Al-Sohaibani S. (2015). Rifampicin resistance among multi-resistant MRSA clinical isolates from Chennai, India, and their molecular characterization. Genetics and molecular research : GMR, 14(1).

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