Quick load taq 2 master mix

Manufactured by New England Biolabs

Sourced in United States

Quick-Load® Taq 2 × Master Mix is a ready-to-use solution that combines Taq DNA Polymerase, dNTPs, and reaction buffer for PCR amplification. It is designed to simplify PCR setup and provide consistent results.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (2 mentions) Primescript rt reagent kit, by Takara Bio (2 mentions) Qiaamp dna mini kit, by Qiagen (1 mentions) Cfx96 real time pcr detection system, by Bio-Rad (1 mentions) Itaq universal sybr green supermix, by Bio-Rad (1 mentions) Abi 3130, by Thermo Fisher Scientific (1 mentions) Mycycler, by Bio-Rad (1 mentions) S1000 thermal cycler, by Bio-Rad (1 mentions) Abi 3730 l, by Thermo Fisher Scientific (1 mentions) Lunascript rt supermix, by New England Biolabs (1 mentions) Qiaamp cador pathogen mini kit, by Qiagen (1 mentions) Gelred, by Biotium (1 mentions)

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Taq Master Mix (17 citations) Quick-Load 2× master mix (2 citations)

7 protocols using quick load taq 2 master mix

Crispr/Cas9 Super-Enhancer Deletion

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To confirm the Crispr/cas9-mediated deletion of super-enhancer peak, genomic DNA was extracted using the QIAamp DNA Mini Kit (Qiagen #51304), followed by PCR to amplify target sequences with or without deletion. Quick-Load® Taq 2× Master Mix (M0270L) was purchased from NEB. Primers flanking the deleted region are designed using Primer-blast online tool and synthesised by Integrated DNA Technologies company (Table S1). PCR products were resolved on a 1.3% agarose gel.

Hu J., Lai Y., Huang H., Ramakrishnan S., Pan Y., Ma V.W., Cheuk W., So G.Y., He Q., Geoffrey Lau C., Zhang L., Cho W.C., Chan K.M., Wang X, & Rebecca Chin Y. (2021). TCOF1 upregulation in triple-negative breast cancer promotes stemness and tumour growth and correlates with poor prognosis. British Journal of Cancer, 126(1), 57-71.

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Comprehensive RNA Extraction and qPCR Analysis

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Total RNA was extracted from cultured cells by using Trizol reagent (Life Technology) and cDNA was generated using PrimeScript RT Reagent kit (Clontech). PCR reactions were carried out by Quick-Load® Taq 2×Master Mix (NEB). PCR was performed under following condition: an initial denaturation at 95 °C for 30 sec.; followed by 28 cycles of 30 sec., denaturation at 95 °C, 20 sec., annealing at 53 °C, and 30 sec. extension at 68 °C; and with a final extension at 68 °C for 10 min.
qPCR was performed using iTaq^TM Universal SYBR Green Supermix (Bio-rad), and detected with the CFX96TM Real-Time PCR Detection System (Bio-rad). qPCR cycling protocol was as follows: an initial denaturation and enzymatic activation at 95 °C for 5 sec; followed by 39 cycles of 5-sec. denaturation at 95 °C, 30-sec. annealing at 60 °C, and melt curve analysis by 0.5 increments at 5 sec./step from 65 °C to 95 °C; and the final incubation was at 95 °C for 30 sec. for polymerase activation and DNA denaturation. Data are normalized to the expressional level of GAPDH and represented as fold changes. Primers used to detect expression of specific genes are listed in Table S2.

Yan L., Jiang B., Li E., Wang X., Ling Q., Zheng D., Park J.W., Chen X., Cheung E., Du X., Li Y., Cheng G., He E, & Xu R.H. (2018). Scalable Generation of Mesenchymal Stem Cells from Human Embryonic Stem Cells in 3D. International Journal of Biological Sciences, 14(10), 1196-1210.

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Genomic DNA Extraction and PCR Amplification

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Cells were pelleted (5 min, 2500 × g, room temperature) and lysed in 400 μL of extraction buffer (200 mM Tris-HCl pH 7.5; 200 mM NaCl; 25 mM EDTA; 0.5% SDS) for 10 min at 37°C under agitation (1400 rpm). After centrifugation (3 min, 17,000 × g, room temperature), the supernatant was harvested and the genomic DNA was precipitated with one volume of isopropanol for 10 min at room temperature, washed with 70% ethanol, dried and resuspended in water. PCR was performed using the Quick-Load^®Taq 2× Master Mix (New England Biolabs) according to the manufacturer recommendations with the primers BSR.5 (5′-GCTGTACGAGGACAACAAGC-3′), TRBCS2.3 (5′-ACGGAGGATCGTTACAACC-3′), CBLP.5 (5′-GACGTCATCCACTGCCTGTG-3′), and CBLP.3 (5′-CGACGCATCCTCAACACACC-3′).

de Carpentier F., Le Peillet J., Boisset N.D., Crozet P., Lemaire S.D, & Danon A. (2020). Blasticidin S Deaminase: A New Efficient Selectable Marker for Chlamydomonas reinhardtii. Frontiers in Plant Science, 11, 242.

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IFNγ-Induced Gene Expression in MSCs

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A total of 10⁵ MSCs were loaded in a 6-well plate and treated with 20 ng/mL IFNγ for 24 h. Total RNA was extracted from the cultured MSCs using Trizol reagent (Life Technology, Carlsbad, CA, USA), and cDNA was generated using the PrimeScript RT Reagent kit (Clontech, Mountain View, CA, USA). PCR reactions were carried out using the Quick-Load^® Taq 2 × Master Mix (NEB). GADPH, indoleamine 2,3-dioxygenase 1 (IDO1), C-C motif chemokine ligand 2 (CCL2), chemokine (C-X-C motif) ligand 10 (CXCL10) and C-X-C motif chemokine receptor 4 (CXCR4) primers were commercially synthesized, and the sequences are listed in Table 1. The PCR conditions are as follows: an initial denaturation at 95 °C for 25 s, followed by 28–32 cycles of 25 s, denaturation at 95 °C for 20 s, annealing at 56 °C for 25 s, extension at 68 °C, and a final extension at 68 °C for 5 min. The PCR product was analyzed by electrophoresis in a 1.5% gel.

Fu X., Jiang B., Zheng B., Yan Y., Wang J., Duan Y., Li S., Yan L., Wang H., Chen B., Sang X., Ji W., Xu R.H, & Si W. (2018). Heterogenic transplantation of bone marrow-derived rhesus macaque mesenchymal stem cells ameliorates liver fibrosis induced by carbon tetrachloride in mouse. PeerJ, 6, e4336.

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COI Amplification and Sanger Sequencing

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Cytochrome oxidase I (COI) PCR products were generated by amplification with LCO1490 and HCO2198 primers (Table 2) (Folmer et al. 1994 (link)). Quick-Load Taq 2× Mastermix (New England Biolabs, Ipswich, MA, USA) was used in PCR reactions with total volumes of 25 µl. Amplification protocols were run on MyCycler or S1000 Thermal Cyclers (BioRad, Hercules, California, USA) for these and all other PCR amplifications. PCR reaction conditions were: 95°C for 5 minutes; 35 cycles of 94°C for 1 minute, 46°C for 1 minute, and 72°C for 1.5 minutes; and a final 5 minute extension at 72°C before being placed on a 4°C hold. PCR products were loaded into a 1% agarose gel in TAE buffer, run for 1 hour at 78 V, and visualized using ethidium bromide under ultraviolet light.
PCR products were sequenced using the Sanger dideoxy method as previously described (Borchers & Marcus 2014 ). PCR products were sequenced in both directions with the primers used to generate the products. Sequencing reactions were analyzed on ABI 3130 or ABI 3730×l automated sequencers (Applied Biosystems, Carlsbad, California, USA) and edited using Sequencher 4.6 software (Sequencher 2005 ). Primer sequences were removed leaving sequenced amplified products of 658 bp, which were then aligned in CLUSTAL W version 2.1 (Thompson et al. 1994 (link); Larkin et al. 2007 (link)).

Gemmell A.P, & Marcus J.M. (2015). A tale of two haplotype groups: Evaluating the New World Junonia ring species hypothesis using the distribution of divergent COI haplotypes. Systematic entomology, 40(3), 532-546.

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Flavivirus Detection in Owl Brain

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Nucleic acids were isolated from the brain of the dead owl by QIAamp^® cador^® Pathogen Mini Kit (Qiagen, Hilden, Germany). The cDNA was synthesized by LunaScript RT SuperMix (New England Biolabs, Ipswich, MA, USA). The presence of flavivirus RNA was examined using a semi-nested PCR (Quick-Load^® Taq 2 × Master Mix, New England Biolabs, Ipswich, MA, USA) by amplification of a partial non-structural (NS) 5 gene. In the first run, PanFlavi-F and cFD2 primers were used to amplify a 599 bp fragment, and in the second run, the primer pair of PanFlavi-F and PanFlavi-R amplified a 360 bp fragment [16 (link),17 (link)]. Amplicons were extracted from an agarose gel (Wizard gel, Promega, Madison, WI, USA) and analyzed by Sanger sequencing (Microsynth, Balgach, Switzerland).

Peňazziová K., Korytár Ľ., Pastorek P., Pistl J., Rusňáková D., Szemes T., Čabanová V., Ličková M., Boršová K., Klempa B, & Csank T. (2021). Genetic Characterization of a Neurovirulent West Nile Virus Variant Associated with a Fatal Great Grey Owl Infection. Viruses, 13(4), 699.

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Genotyping Mouse Ear Snips by PCR

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DNA samples were prepared from mouse ear snips using NaOH extraction, and analyzed by PCR. PCR was performed using Quick-Load Taq 2× Master Mix (New England Biolabs). PCR conditions: denaturation at 94°C for 5 min followed by 30× 94°C for 30 s, 58°C for 30 s, 68°C for 45 s, 68°C for 5 min. Primers specific for Cre allele, internal control, floxed NR1 alleles were according to reference [25] (link). PCR products were analyzed by electrophoresis on 2% agarose with GelRed (Biotium).

Pozzi L., Dorocic I.P., Wang X., Carlén M, & Meletis K. (2014). Mice Lacking NMDA Receptors in Parvalbumin Neurons Display Normal Depression-Related Behavior and Response to Antidepressant Action of NMDAR Antagonists. PLoS ONE, 9(1), e83879.

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