Taq dna polymerase 2 master mix

Manufactured by Ampliqon

Sourced in Denmark

Taq DNA Polymerase 2 × Master Mix is a ready-to-use, pre-mixed solution containing Taq DNA polymerase, dNTPs, and reaction buffer. It is designed for high-throughput and routine PCR applications.

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Lab products found in correlation

T100 thermal cycler, by Bio-Rad (1 mentions) Taqman genotyping assay, by Thermo Fisher Scientific (1 mentions) Lightcycler 96 system, by Roche (1 mentions) Thermocycler, by Thermo Fisher Scientific (1 mentions) Mgcl2, by Ampliqon (1 mentions) Gel documentation system, by Uvitec (1 mentions)

Close spelling variants of this product

Taq DNA polymerase (23 citations) Taq DNA Polymerase 2× Master Mix RED (16 citations) Taq polymerase (6 citations) 2× Taq master Mix (4 citations) Taq DNA polymerase 2.0× Master Mix RED (3 citations)

4 protocols using taq dna polymerase 2 master mix

Sequence Analysis of vra Genes in S. aureus

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WalKR, VraTSR, and GraSR are the main transcriptional regulator systems in S. aureus. The vraS and vraT genes were selected for further sequence analysis due to modification in their gene expression levels (see the results of the RT-qPCR analysis).
Genomic DNA extraction was performed according to the Gene Transfer kit (Pioneers, Iran) instructions. The complete sequence of vraS and vraT genes were amplified by PCR in a T100™ Thermal cycler (BioRad, USA) using Taq DNA Polymerase 2 × Master Mix (Ampliqon, Denmark) and previously published primers [3 (link)].
After verifying using gel electrophoresis, the PCR products were purified and sequenced by Microsynth AG company (Switzerland).
The sequence read of CHG-induced CHG_Van-I strain was compared to wild-type VAN-S strain using the alignment in allele ID software (V6.00, USA) and BLAST database (https://blast.ncbi.nlm.nih.gov/Blast.cgi) to find CHG-induced single nucleotide polymorphisms (SNPs).

Baseri N., Najar-Peerayeh S., Bakhshi B, & Campanile F. (2022). Phenotypic and genotypic changes of Staphylococcus aureus in the presence of the inappropriate concentration of chlorhexidine gluconate. BMC Microbiology, 22, 100.

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Genotyping of Selected Genetic Variants

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Genotyping of selected variants was performed by TaqMan Genotyping Assays, Applied Biosystems (Assay ID: C__3127459_10, Assay ID: C__26124318_10, Assay ID: C__1085366_10). The 2X master mix was purchased from Ampliqon (Taq DNA Polymerase 2× Master Mix). The reaction was performed in 10 μl volume, based on the following formula of 5 μl of 2× master mix, 0.3 μl of the primer‐probe mixture, 2 μl of DNA, and 2.7 μl of nuclease‐free water. The reaction program was 10 min at 60°C (pre‐incubation phase) and production cycles contained 10 s 95°C and 40 s 60°C for each of 40 cycles. TaqMan genotyping polymerase chain reactions (PCR) were performed in Roche LightCycler® 96 System and analyzed by LightCycler 96sw1.1 software. To confirm the accuracy of the TaqMan method, for each variant, three samples of heterozygote, wild type homozygote, and mutant homozygote had repeated TaqMan reaction. To ensure the absence of any possible contamination in the reactions, a well was assigned as the blank control without adding the DNA.

Gholami M., Zoughi M., Behboo R., Taslimi R., Kazemeini A., Bastami M., Hasani‐Ranjbar S., Larijani B, & Amoli M.M. (2022). Association of miRNA targetome variants in LAMC1 and GNB3 genes with colorectal cancer and obesity. Cancer Medicine, 11(21), 3923-3938.

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Genotyping CARTPT rs2239670 Polymorphism

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The genomic DNA of participants was extracted from the whole blood using a standard phenol/chloroform method. Subjects were genotyped for the CARTPT rs2239670 (major allele: G; minor allele: A) polymorphism using polymerase chain reaction-restricted length polymorphism (PCR–RFLP) technique. Template primers used for the PCR amplification of the rs2239670 were as follow: forward: CCTGCTGCTGATGCTACCTCT-3′ and reverse: 5′-GCGCTTCGATCTGCAACACAC-3′. The PCR reaction was optimized in a total volume of 25 μl containing 0.5 μl of each primer, 2 μl of DNA, 10 μl of Taq DNA Polymerase 2 × MasterMix (Ampliqon, Denmark) and 12.5 μl distilled water. For PCR amplification, the following conditions were applied: 94 °C/5 min (initial denaturation), 35 cycles of denaturation 94 °C/30 s, annealing 60 °C/30 s, extension 72 °C/20 s and 72 °C/10 min (final synthesis). Digestion of the amplified DNA was performed with ApaI (Takara, Japan) restriction enzyme overnight. The digested products were then analyzed by electrophoresis on 3% agarose gel. Homozygous for wild-type allele (GG) of the CARTPT rs2239670 was distinguished as cut fragments (340 and 212 bp), heterozygous as cut fragments (212, 340 and 552 bp) and homozygous for mutant allele (AA) as uncut fragment (552 bp).

Khodarahmi M., Siri G., Erahimzadeh F., Farhangi M.A, & Shanehbandi D. (2022). Dietary glycemic index and glycemic load mediate the effect of CARTPT rs2239670 gene polymorphism on metabolic syndrome and metabolic risk factors among adults with obesity. BMC Endocrine Disorders, 22, 288.

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Detection of ESBL-Associated Genes by PCR

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ESBL-producing isolates were subjected to polymerase chain reaction (PCR) for detection of bla OXA , bla PER-1 bla VEB-1 , and bla GES-1 genes using the specific primers listed in Table 1 [18] [19] [20] [21] [22] [23] [24] . Total deoxyribonucleic acid (DNA) was extracted from bacterial isolates using an extraction kit (Bioneer, Daejeon, Korea). PCR amplification was performed in a thermocycler (Applied Biosystems, Carlsbad, CA, USA) as follows: 96 o C for 5 min; 35 cycles of 1 min at 96 o C, 1 min at a specific temperature for each primer, and 1 min at 72 o C; and a final extension step of 10 min at 72 o C. Amplification reactions were prepared in a total volume of 25μl including 12.5μl Taq DNA polymerase 2× Master Mix with 1.5mM MgCl 2 (Ampliqon, Odense, Denmark), 0.5μM forward primer, 0.5μM reverse primer, 9μl nuclease free water, and 2.5μl DNA template (50pg concentration). PCR products were electrophoresed on a 1% agarose gel at 100V, stained with ethidium bromide solution, and finally visualized with a gel documentation system (UviTec, Cambridge, UK). Purified PCR products were sequenced by the Macrogen Company (Seoul, Korea), and sequence alignment and analysis were performed online using the basic local alignment search tool (BLAST) program of the National Center for Biotechnology Information (http://blast.ncbi.nlm.nih.gov/Blast.cgi).

Amirkamali S., Naserpour-Farivar T., Azarhoosh K, & Peymani A. (2017). Distribution of the bla OXA , bla VEB-1 , and bla GES-1 genes and resistance patterns of ESBL-producing Pseudomonas aeruginosa isolated from hospitals in Tehran and Qazvin, Iran. Revista da Sociedade Brasileira de Medicina Tropical, 50(3).

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