Taq reaction buffer

Manufactured by New England Biolabs

Sourced in United Kingdom

Taq reaction buffer is a specialized buffer solution designed to support the activity of Taq DNA polymerase, a thermostable enzyme commonly used in the polymerase chain reaction (PCR) process. It provides the necessary ions and pH conditions to facilitate the efficient amplification of DNA sequences.

Automatically generated - may contain errors

Lab products found in correlation

Taq polymerase, by New England Biolabs (3 mentions) Dntps, by Thermo Fisher Scientific (2 mentions) C1000 thermal cycler, by Bio-Rad (1 mentions) Proteinase k, by Merck Group (1 mentions) Nupage 12 bis tris gel, by Thermo Fisher Scientific (1 mentions) Bigdye v3, by Thermo Fisher Scientific (1 mentions) Dneasy blood and tissue kit, by Qiagen (1 mentions)

Close spelling variants of this product

Standard Taq Reaction Buffer LongAmp Taq Reaction Buffer × Taq DNA ligase reaction buffer HiFi Taq DNA Ligase Reaction Buffer Reaction buffer Taq buffer

5 protocols using taq reaction buffer

Mitochondrial DNA Purity Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The purity of the extracted mtDNA was analyzed by PCR amplification using mitochondrial, COX1; nuclear, actin; chloroplast, RBCL genes specific primers (see Additional file 1). PCRs were performed on C1000 Thermal Cycler (Bio-Rad) in 20 µl volumes containing 120 ng mtDNA, 2 µl of 10X standard Taq reaction buffer (New England Bio Labs, cat. no. B9014S, Whitby, ON, Canada), 0.1 mM dNTPs, 0.25 pM primers (forward and reverse each), 0.3 µl of Taq polymerase (New England Bio Labs, cat. no. M0273X, Whitby, ON, Canada) and ddH₂O to make volume to 20 µl. Reaction conditions were as follow: 95 °C for 5 min followed by 30 cycles at 95 °C for 30 s, 55 °C (actin) or 60 °C (COX1, RBCL) for 30 s and 72 °C for 1 min and a final extension at 72 °C for 7 min.
To visualize PCR products for purity and quality, agarose gel electrophoresis was applied. Approximately 28 µg of purified mtDNA or PCR reaction from each sample were loaded onto 1.5 % agarose gel. The electrophoresis was carried out in 1x TAE buffer (40 mM Tris-Acetate, 1 mM EDTA) at 100 V and 3 mAh for 3 h. The gels were visualized by staining with 0.5 mg/mL ethidium bromide.

Ahmed Z, & Fu Y.B. (2015). An improved method with a wider applicability to isolate plant mitochondria for mtDNA extraction. Plant Methods, 11, 56.

+ Open protocol

+ Expand

Quantifying mtDNA Content in Worms

Check if the same lab product or an alternative is used in the 5 most similar protocols

mtDNA content relative to nuclear DNA in worms was quantified using a previously described method (53 ). Briefly, age-synchronized young adult worms grown on NGM plates at 25°C were added to lysis buffer [1× standard Taq reaction buffer (New England Biolabs) containing proteinase K (1 mg/ml; Sigma-Aldrich)] in batches of eight worms/60 μl of buffer and were immediately frozen at −80°C for 15 min. Samples were incubated at 65°C for 1 hour, followed by at 95°C for 15 min. Mitochondrial and nuclear DNA content were estimated using real-time quantitative PCR, as described previously, with PCR primers listed in data file S2. Ratio of mtDNA:nuclear DNA copy number was calculated using comparative Ct method.

Sural S., Liang C.Y., Wang F.Y., Ching T.T, & Hsu A.L. (2020). HSB-1/HSF-1 pathway modulates histone H4 in mitochondria to control mtDNA transcription and longevity. Science Advances, 6(43), eaaz4452.

+ Open protocol

+ Expand

Taq Polymerase Stalling at Abasic Sites

Check if the same lab product or an alternative is used in the 5 most similar protocols

To confirm that Taq polymerase permanently stalls at abasic sites (and by extension, lesions attached to abasic sites) 200 pmol of the oligodeoxynucleotide M13-RSV-Zeo-8oxo (template) was annealed to 200 pmol of primer C (Table 1) in restriction enzyme buffer 2 (New England Biolabs) and water in a volume of 10 μL at 95°C for 5 minutes and slowly cooled to room temperature. The creation of a duplex is necessary because OGG1 protein will not react with single-stranded DNA. The duplex DNA was then incubated in the presence or absence of OGG1 (200 pmol) in buffer containing 100 mM NaCl, 1mM MgCl₂, and 20 mM Tris-HCl pH 7.0 at 37°C (final volume 15 μL) for 30 minutes. Samples incubated with OGG1 excise the 8-oxo-dG residue leaving an abasic site at its position. Each sample was divided in half and incubated in the presence or absence of primer Z (100 pmol, Table 1) with 10 mM dNTPs (Thermo Scientific), Taq reaction buffer (New England Biolabs), and Taq polymerase (20 units) in a final volume of 10 μL at 95°C for 5 minutes. Samples were then slow cooled to 75°C for 15 minutes, resolved on a NuPAGE 12% Bis-Tris Gel (Invitrogen) at 18V/cm at 55°C, and stained with ethidium bromide.

Chesner L.N, & Campbell C. (2018). A quantitative PCR-based assay reveals that nucleotide excision repair plays a predominant role in the removal of DNA-protein crosslinks from plasmids transfected into mammalian cells. DNA repair, 62, 18-27.

+ Open protocol

+ Expand

Genomic DNA extraction and SCN4A sequencing

Check if the same lab product or an alternative is used in the 5 most similar protocols

We extracted whole genomic DNA from ethanol-preserved liver samples using a DNeasy Blood and Tissue Kit (Qiagen) following the manufacturer’s protocol. We PCR-amplified the Domain I Subunit 6 (DI-S6) of the Na_V1.4 sodium ion channel encoded by the SCN4A gene. We targeted this locus using primers +869F (5’-CTGCAGGYAAAACCTACATGG-3’;) and +1032R (5’-GATGTTTCTTTAGCTGTTCC-3’) [38 (link)]. For all samples, we conducted 25μL reactions using 2% dimethyl sulfoxide, 1X standard Taq reaction buffer (NEB), 1.5mM MgCl₂, 0.2mM dNTPs, 0.2μM final concentrations for each primer, and 0.625U Taq polymerase (NEB). We ran PCRs at an initial temperature of 94°C for 2min, followed by 30 cycles of 94°C for 30s, 51°C for 30s, and 72°C for 30s, followed by a final extension at 72°C for 5min. We then purified amplicons using ExoSAP-IT (USB). Next, we performed cycle sequencing reactions using Big Dye v3.1 sequencing chemistry (Applied Biosystems) and sequenced Sephadex^™ G-50 cleaned products on an ABI Automated 3730xl DNA Analyzer (Applied Biosystems). We examined electropherograms by eye and assembled sequences in Sequencher v4.8. Finally, we deposited all newly generated sequences in EMBL-ENA (accession numbers: LT984926-LT984986).

Yuan M.L, & Wang I.J. (2018). Sodium ion channel alkaloid resistance does not vary with toxicity in aposematic Dendrobates poison frogs: An examination of correlated trait evolution. PLoS ONE, 13(3), e0194265.

+ Open protocol

+ Expand

Multiplex PCR for ExPEC Virulence Genes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Eight virulence genes, viz. traT, ompT, iutA, fimH, hlyA, iha, papC, and cvaC, which are commonly associated with ExPEC were characterized by multiplex PCR methods. Three positive control genes (fimH, hlyA, and iutA) were synthesized and commercially cloned in GeneArt vectors (Invitrogen, USA). The recombinant plasmids were then transformed into a DH5α competent cell using Subcloning Efficiency™ DH5α Competent Cell (Invitrogen, USA). The primers for these three virulence genes (designed here) and five primers sets for traT, ompT, cvaC, iha, and papC genes (described previously) are tabulated in Supplementary Table S4. The first pool consists of fimH, hlyA, and iutA, while the second pool comprises cvaC, iha, traT, ompT, and papC. A total 25 µL reaction comprising 1 × Standard Taq reaction buffer and 1.5 U Taq polymerase (New England Biolabs, UK), 0.2 µM of each set of primers (forward and reverse), 0.2 mM dNTPs, ~ 100 ng DNA template and ultra-pure water were used. The optimized PCR condition for virulence genes was: initial denaturation at 94 ℃ for 15 min followed by 29 cycles at 94 ℃ for 1 min, annealing at 56 ℃ for 1 min, extension at 72 ℃ for 1 min, and post extension at 72 ℃ for 10 min. All amplicons were analyzed using 2% agarose gel electrophoresis.

Ullah N., Assawakongkarat T., Akeda Y, & Chaichanawongsaroj N. (2023). Detection of Extended-spectrum β-lactamase-producing Escherichia coli isolates by isothermal amplification and association of their virulence genes and phylogroups with extraintestinal infection. Scientific Reports, 13, 12022.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!