standard taq reaction buffer  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Name:
    Standard Taq Reaction Buffer Pack
    Description:
    Standard Taq Reaction Buffer Pack 6 0 ml
    Catalog Number:
    B9014S
    Price:
    22
    Size:
    6 0 ml
    Category:
    Buffers
    Score:
    85
    Buy from Supplier


    Structured Review

    New England Biolabs standard taq reaction buffer
    Standard Taq Reaction Buffer Pack
    Standard Taq Reaction Buffer Pack 6 0 ml
    https://www.bioz.com/result/standard taq reaction buffer/product/New England Biolabs
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    standard taq reaction buffer - by Bioz Stars, 2019-11
    99/100 stars

    Images

    Related Articles

    Amplification:

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr. .. The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr.

    Article Title: Impact of swapping soils on the endophytic bacterial communities of pre-domesticated, ancient and modern maize
    Article Snippet: A PCR mastermix was made with the following components per 25 μl volume: 2.5 μl Standard Taq Buffer (New England Biolabs), 0.5 μl of 25 mM dNTP mix, 0.5 μl of 10 mM 27 F-Degen primer with sequence AGRRTTYGATYMTGGYTYAG [ ] (where R = A + G, Y = C + T, M = A + C), 0.5 μl of 10 mM 1492r primer with sequence GGTTACCTTGTTACGACTT [ ], 0.25 μl of 50 mM MgCl2 , 0.25 μl of Standard Taq (New England Biolabs), 50 ng of total DNA, and double distilled water up to 25 μl total. .. A PCR mastermix was made with the following components per 25 μl volume: 2.5 μl Standard Taq Buffer (New England Biolabs), 0.5 μl of 25 mM dNTP mix, 0.5 μl of 10 mM 27 F-Degen primer with sequence AGRRTTYGATYMTGGYTYAG [ ] (where R = A + G, Y = C + T, M = A + C), 0.5 μl of 10 mM 1492r primer with sequence GGTTACCTTGTTACGACTT [ ], 0.25 μl of 50 mM MgCl2 , 0.25 μl of Standard Taq (New England Biolabs), 50 ng of total DNA, and double distilled water up to 25 μl total.

    Article Title: Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments
    Article Snippet: When the plants reached the 1 to 2 leaf stage, 3 cm leaf segments from primary leaves were harvested for genomic DNA isolation. .. A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process. .. The PCR conditions were: initial denaturation at 94 °C (3 min), followed by 44 cycles of 94 °C (1 min), 55 or 60 °C annealing (1 min), and 72 °C extension (1 min) with a final extension at 72 °C for 10 min.

    Article Title: Ocean currents influence the genetic structure of an intertidal mollusc in southeastern Australia - implications for predicting the movement of passive dispersers across a marine biogeographic barrier
    Article Snippet: Approximately 650 base pairs (bp) of the mitochondrial cytochrome oxidase I gene (COI ) was amplified for 10 individuals from each sample site by PCR using primers HCO and LCO (Folmer et al. ). .. PCRs were prepared in 25 μL volumes each containing 10.25 μL ddH2O, 1× reaction buffer, 5.0 μL dNTPs (1 mmol/L), 0.4 μmol/L HCO primer (10 μmol/L), 0.4 μmol/L LCO primer (10 μmol/L), 0.25 units NEB taq (Biosciences, New England), and 5 μL DNA extract.

    Article Title: CRISPR-MVLST subtyping of Salmonella enterica subsp. enterica serovars Typhimurium and Heidelberg and application in identifying outbreak isolates
    Article Snippet: Paragraph title: PCR amplification ... PCR reactions were performed in a total volume of 25 μl: 1.5 μl template, 0.3 μl Taq (1.5 units; New England Bio Labs, Ipswich, MA), 0.2 μl 10 mM dNTPs, 1 μl of each 10 μM primer, 2.5 μl of 10× Taq buffer and 18.5 μl water.

    Article Title: RAD marker microarrays enable rapid mapping of zebrafish mutations
    Article Snippet: PCR reactions were carried out in 50 μl under the following conditions: 50 ng template, 1 × Thermopol Buffer (New England Biolabs), 2.5 U Taq (New England Biolabs), 0.2 mmol/l dNTPs (Fermentas, Burlington, Ontario, Canada), 0.4 μmol/l forward primer, and 0.2 μmol/l reverse primer. .. Touchdown amplification was applied, with primer annealing temperature dropping 2°C every other cycle from 65°C to 55°C, after which 25 additional cycles were carried out [ ].

    Article Title: A Simple Isothermal DNA Amplification Method to Screen Black Flies for Onchocerca volvulus Infection
    Article Snippet: LAMP primers B3 and F3 were used to PCR amplify OvGST1a in 25 µL reactions containing 3 µL DNA template, 0.2 µM of each primer, and 1.25 U of Taq DNA polymerase in 1× standard buffer (New England Biolabs) containing 3.5 mM MgCl2 , 0.2 mM and 0.2 mM dNTP each. .. PCR products were visualized by UV transillumination in a 1.5% agarose gel after electrophoresis and staining with ethidium bromide.

    Article Title: 16S rRNA Terminal Restriction Fragment Length Polymorphism for the Characterization of the Nasopharyngeal Microbiota
    Article Snippet: Purified PCR products were checked for correct amplification size and purity using the Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, CA) with the DNA 7500 LabChip kit. .. The enzymatic digestions were performed in a total volume of 20 µl using 40 ng of the amplified and purified PCR product, 1 µl NEB Reaction Buffer (New England Biolabs, Ipswich, MA) 1 (for digestion with Tsp509I ) or 4 (Hpy166II ) and 0.5 µl of the restriction enzymes Tsp509I or Hpy166II . .. Digestion conditions were 3 h at 65°C for TSP509I and 3 h at 37°C with 20 min enzyme inactivation at 65°C for Hpy166II , respectively.

    Positive Control:

    Article Title: A Simple Isothermal DNA Amplification Method to Screen Black Flies for Onchocerca volvulus Infection
    Article Snippet: LAMP primers B3 and F3 were used to PCR amplify OvGST1a in 25 µL reactions containing 3 µL DNA template, 0.2 µM of each primer, and 1.25 U of Taq DNA polymerase in 1× standard buffer (New England Biolabs) containing 3.5 mM MgCl2 , 0.2 mM and 0.2 mM dNTP each. .. PCR products were visualized by UV transillumination in a 1.5% agarose gel after electrophoresis and staining with ethidium bromide.

    Synthesized:

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: First strand cDNA was synthesized using ProtoScript M-MuLV FS-cDNA Synthesis Kit (New England Biolabs, MA) according to the manufacturer’s protocol and modified oligonucleotides in Supporting Information , Table S1 . .. The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr.

    Neutralization:

    Article Title: Vitamin E inhibits the UVAI induction of “light” and “dark” cyclobutane pyrimidine dimers, and oxidatively generated DNA damage, in keratinocytes
    Article Snippet: T4endoV (0.1 U/mL), or hOGG1 (3.2 U/mL), or enzyme reaction buffer alone, was added to each gel (both enzymes were purchased from New England Biolabs, Hitchin, UK). .. Slides were subsequently transferred to ice-cold electrophoresis buffer (NaOH 10 M, EDTA 200 mM, pH 13 in ddH2 O) and incubated for 20 min in the dark.

    Terminal Restriction Fragment Length Polymorphism:

    Article Title: Impact of swapping soils on the endophytic bacterial communities of pre-domesticated, ancient and modern maize
    Article Snippet: Paragraph title: DNA extraction and Terminal Restriction Fragment Length Polymorphism (TRFLP) from plant tissues and soil ... A PCR mastermix was made with the following components per 25 μl volume: 2.5 μl Standard Taq Buffer (New England Biolabs), 0.5 μl of 25 mM dNTP mix, 0.5 μl of 10 mM 27 F-Degen primer with sequence AGRRTTYGATYMTGGYTYAG [ ] (where R = A + G, Y = C + T, M = A + C), 0.5 μl of 10 mM 1492r primer with sequence GGTTACCTTGTTACGACTT [ ], 0.25 μl of 50 mM MgCl2 , 0.25 μl of Standard Taq (New England Biolabs), 50 ng of total DNA, and double distilled water up to 25 μl total.

    Article Title: 16S rRNA Terminal Restriction Fragment Length Polymorphism for the Characterization of the Nasopharyngeal Microbiota
    Article Snippet: Paragraph title: T-RFLP Digestion and Detection Limits ... The enzymatic digestions were performed in a total volume of 20 µl using 40 ng of the amplified and purified PCR product, 1 µl NEB Reaction Buffer (New England Biolabs, Ipswich, MA) 1 (for digestion with Tsp509I ) or 4 (Hpy166II ) and 0.5 µl of the restriction enzymes Tsp509I or Hpy166II .

    SYBR Green Assay:

    Article Title: Prp8 retinitis pigmentosa mutants cause defects in the transition between the catalytic steps of splicing
    Article Snippet: Reactions were then diluted between 1:75 and 1:250 with water prior to analysis by qPCR. .. For qPCR, 10 µL of diluted cDNA from reverse transcription was combined with 0.25 mM each gene-specific primer, 1× SYBR green (Roche), 0.625 µL dimethyl sulfoxide (Invitrogen), 0.4 mM dTNPs (Fermentas), 1× Standard Taq Buffer (NEB) and 0.25 µL Standard Taq (NEB) and water to a total volume of 25 µL. qPCR cycle conditions were 95°C 3 min, 39 cycles of 95°C 15 sec, 55°C 30 sec, 72°C 15 sec, followed by one cycle at 72°C for 3 min. Exon primers were designed to be within the 3′ exon of all genes tested and were used to measure total RNA levels. pre-mRNA levels were assessed using intron primers. .. The product of these primes spanned an intron–exon junction to ensure that only pre-mRNA was measured.

    Article Title: Cooperative activity of GABP with PU.1 or C/EBPε regulates lamin B receptor gene expression, implicating their roles in granulocyte nuclear maturation
    Article Snippet: Real-time PCR was performed by using 2 μl of purified DNA as template, SsoAdvanced SYBR Green Supermix (Bio-Rad) and the following primers at a final concentration of 0.2 pmol/μl: Proximal Ets site: (F) 5′-AACAGCACGGCGAAGTAG-3′ and (R) 5′-CCCCTTACACTTGCCACC-3′, Distal Ets site: (F) 5′-GATCTGTTTGTCCCCAGCTAC-3′ and (R) 5′-ACTTTGTGAGCACTTGAGACC-3′, and Proximal Cebp site: (F) 5′-CTCGGAGTAGGATTCGTCTTTAAG-3′ and (R) 5′-GATTTGTCATTGCCGTTGGG-3′. .. A 25 μl PCR reaction was also performed using 2 μl of purified DNA from EML cells using 1X standard Taq reaction buffer (New England BioLabs), 1 U of Taq DNA polymerase, plus 0.2 pmol/μl of same forward and reverse primers for proximal and distal Ets sites as mentioned above.

    Incubation:

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr. .. Tailed templates were ligated to double stranded adaptors prepared with oligonucleotides from the Illumina Customer Sequence Letter (version August 12, 2014, ; Table S1 ).

    Article Title: Prp8 retinitis pigmentosa mutants cause defects in the transition between the catalytic steps of splicing
    Article Snippet: Briefly, for reverse transcription 2 µg of DNase-treated total RNA was incubated with 0.5 µL of 1 mg/mL dN9 primers (Life Technologies) in a total volume of 10 µL at 65°C for 5 min and then chilled on ice for 5 min. Four microliters of 5× First Strand Synthesis Buffer (Invitrogen), 1 µL 10 mM dNTPs (Fermentas), 1 µL 0.1 M DTT (Invitrogen), 1 µL Superscript III Reverse Transcriptase (Invitrogen), and 3 µL of water were added to bring the total reaction volume to 20 µL. .. For qPCR, 10 µL of diluted cDNA from reverse transcription was combined with 0.25 mM each gene-specific primer, 1× SYBR green (Roche), 0.625 µL dimethyl sulfoxide (Invitrogen), 0.4 mM dTNPs (Fermentas), 1× Standard Taq Buffer (NEB) and 0.25 µL Standard Taq (NEB) and water to a total volume of 25 µL. qPCR cycle conditions were 95°C 3 min, 39 cycles of 95°C 15 sec, 55°C 30 sec, 72°C 15 sec, followed by one cycle at 72°C for 3 min. Exon primers were designed to be within the 3′ exon of all genes tested and were used to measure total RNA levels. pre-mRNA levels were assessed using intron primers.

    Article Title: Vitamin E inhibits the UVAI induction of “light” and “dark” cyclobutane pyrimidine dimers, and oxidatively generated DNA damage, in keratinocytes
    Article Snippet: T4endoV (0.1 U/mL), or hOGG1 (3.2 U/mL), or enzyme reaction buffer alone, was added to each gel (both enzymes were purchased from New England Biolabs, Hitchin, UK). .. T4endoV (0.1 U/mL), or hOGG1 (3.2 U/mL), or enzyme reaction buffer alone, was added to each gel (both enzymes were purchased from New England Biolabs, Hitchin, UK).

    Article Title: Protein-Tyrosine Kinase Activity Profiling in Knock Down Zebrafish Embryos
    Article Snippet: The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array. .. The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array.

    Activity Assay:

    Article Title: Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments
    Article Snippet: When the plants reached the 1 to 2 leaf stage, 3 cm leaf segments from primary leaves were harvested for genomic DNA isolation. .. A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process. .. The PCR conditions were: initial denaturation at 94 °C (3 min), followed by 44 cycles of 94 °C (1 min), 55 or 60 °C annealing (1 min), and 72 °C extension (1 min) with a final extension at 72 °C for 10 min.

    In Silico:

    Article Title: Impact of swapping soils on the endophytic bacterial communities of pre-domesticated, ancient and modern maize
    Article Snippet: A PCR mastermix was made with the following components per 25 μl volume: 2.5 μl Standard Taq Buffer (New England Biolabs), 0.5 μl of 25 mM dNTP mix, 0.5 μl of 10 mM 27 F-Degen primer with sequence AGRRTTYGATYMTGGYTYAG [ ] (where R = A + G, Y = C + T, M = A + C), 0.5 μl of 10 mM 1492r primer with sequence GGTTACCTTGTTACGACTT [ ], 0.25 μl of 50 mM MgCl2 , 0.25 μl of Standard Taq (New England Biolabs), 50 ng of total DNA, and double distilled water up to 25 μl total. .. For the nested PCR, primer 799f with sequence AACMGGATTAGATACCCKG [ ] (where M = A + C, K = G + T), was labelled with 6FAM, and 1492rh with sequence HGGHTACCTTGTTACGACTT (where H = A + T + C) was labelled with Max550, both by Integrated DNA Technologies (USA).

    Modification:

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: First strand cDNA was synthesized using ProtoScript M-MuLV FS-cDNA Synthesis Kit (New England Biolabs, MA) according to the manufacturer’s protocol and modified oligonucleotides in Supporting Information , Table S1 . .. The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr.

    Article Title: Real-Time PCR Assays To Address Genetic Diversity among Strains of Mycoplasma hyopneumoniae
    Article Snippet: These assays were then used to test DNA from each of the isolates in Table . .. Some assays were performed as previously published ( , , ), while others ( , , , , ) were modified by substituting MasterAmp E buffer (Epicentre Biotechnologies, Madison, WI) for the published buffer with their published cycling parameters in reaction mixtures consisting of 1× Master Amp E, 0.4 mM each forward and reverse primer, 2.5 U of Taq (New England Biolabs, Inc., Ipswich, ME), template containing 1 to 10 ng of DNA, and a sufficient quantity of PCR-grade water to a final volume of 25 μl. .. The primer pairs from nested-set PCRs were run as individual assays (not as nested sets) to independently evaluate the ability of each primer pair to form products.

    Derivative Assay:

    Article Title: Protein-Tyrosine Kinase Activity Profiling in Knock Down Zebrafish Embryos
    Article Snippet: Each peptide represents a 15 amino acid sequence, of which 13 residues are derived from a known phosphorylation site from Swissprot and Phosphobase databases ( ). .. The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array.

    Ligation:

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr. .. Purified, tailed cDNA was combined with T4 DNA ligase buffer (New England Biolabs, MA), T4 DNA ligase (New England Biolabs, MA), and the double stranded adaptors, and the solution was incubated at 12° for at least 6 hr.

    Infection:

    Article Title: A new insect cell glycoengineering approach provides baculovirus-inducible glycogene expression and increases human-type glycosylation efficiency
    Article Snippet: At various times after infection with AchEPO-His, total RNA was extracted from 5×106 Sf9, Sfie1SWT, or Sf39KSWT cells using the RNA- Solv ® (Omega Biotech) reagent according to the manufacturer’s protocol. .. The resulting cDNA preparations were treated with RNAse H and used for PCR reactions with Taq Polymerase and Crimson Taq Buffer (New England Biolabs).

    Generated:

    Article Title: Allopatric Speciation within a Cryptic Species Complex of Australasian Octopuses
    Article Snippet: 25 µL reactions comprised 0.1 µL Taq (Onetaq, New England Biolabs ), 2.5 µL 10 x buffer (Paq5000™), 2 µL dNTP mix (10 µM, Bioline ), 0.5 µL forward primer (10 µM), 0.5 µL reverse primer (10 µM), 17.4 µL ddH2 O and 2 µL DNA (diluted to between 1–5 ng/µL). .. 25 µL reactions comprised 0.1 µL Taq (Onetaq, New England Biolabs ), 2.5 µL 10 x buffer (Paq5000™), 2 µL dNTP mix (10 µM, Bioline ), 0.5 µL forward primer (10 µM), 0.5 µL reverse primer (10 µM), 17.4 µL ddH2 O and 2 µL DNA (diluted to between 1–5 ng/µL).

    Polymerase Chain Reaction:

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: Additionally, 2 µL of each reaction was separated on a 1% agarose gel containing GelRed to ensure successful reactions. .. The barcoding PCR reaction consisted of 0.235 µl of H2 O, 0.625 µl NEB Standard Taq Buffer (1.25 ×), 0.1 µl dNTPs (500 µM), 1 µl universal reverse primer (1 µM), 1 µl unique barcode adaptor primer (0.4 µM), 0.04 µl of NEB Taq polymerase (0.2 units), and 2 µl template (diluted primary PCR reaction). .. PCR parameters are the same as the primary reaction.

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: The parameters for the PCR reactions were as follows: 94° for 5 min, followed by 35 cycles of 94° for 30 sec, 62° for 30 sec, and 72° for 60 sec, followed by a final extension at 72° for 7 min. .. The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions. .. PCR products were visualized on a 1% agarose gel containing GelRed (Biotium, CA) and subsequently purified using E.Z.N.A.

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: Second strand synthesis was performed by incubating first-strand cDNA with 1× NEBNext Second Strand Synthesis Buffer (New England Biolabs, MA), 0.2 mM dNTPs, 15 units of Escherichia coli DNA ligase (New England Biolabs, MA), 75 units of E. coli DNA polymerase I (New England Biolabs, MA), and 3 units of RNase H (New England Biolabs, MA) for 2 hr at 16°. cDNA was purified using the GeneJet PCR Purification Kit (Fermentas, MA) and then fragmented using NEBNext dsDNA Fragmentase (New England Biolabs, MA) according to the manufacturer’s protocol, with the addition of 5 mM MgCl2 and 1 mg ml−1 BSA (New England Biolabs, MA). .. The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr.

    Article Title: Impact of swapping soils on the endophytic bacterial communities of pre-domesticated, ancient and modern maize
    Article Snippet: Total DNA from 250 mg of soil or sand was extracted using Powersoil DNA isolation kits (MoBio, USA), and DNA was quantified using a Nanodrop (Thermo Scientific, USA). .. A PCR mastermix was made with the following components per 25 μl volume: 2.5 μl Standard Taq Buffer (New England Biolabs), 0.5 μl of 25 mM dNTP mix, 0.5 μl of 10 mM 27 F-Degen primer with sequence AGRRTTYGATYMTGGYTYAG [ ] (where R = A + G, Y = C + T, M = A + C), 0.5 μl of 10 mM 1492r primer with sequence GGTTACCTTGTTACGACTT [ ], 0.25 μl of 50 mM MgCl2 , 0.25 μl of Standard Taq (New England Biolabs), 50 ng of total DNA, and double distilled water up to 25 μl total. .. Amplification was for 35 cycles in a PTC200 DNA Thermal Cycler (MJ Scientific, USA) using the following program: 96°C for 3 min, 35X (94°C for 30 sec, 48°C for 30 sec, 72°C for 1:30 min), 72°C for 7 min.

    Article Title: A Recurrent Stop-Codon Mutation in Succinate Dehydrogenase Subunit B Gene in Normal Peripheral Blood and Childhood T-Cell Acute Leukemia
    Article Snippet: The 30th (last) cycle of the PCR was completed to produce fluorescently-labeled homo-duplex extension products. .. Fifteen µL of the fluorescently-labeled PCR products, which contained ∼200–300 ng of DNA, were directly digested by 10 U of Taq I RE, 1× Taq I RE buffer and 1× Bovine Serum Albumin (all from New England Biolabs) overnight in a 25 µL reaction volume. .. To ensure complete digestion, the reaction was re-dosed with another 10 U of Taq I RE in a 5 µL volume and continued for at least another four hours in the following day.

    Article Title: Real-Time PCR Assays To Address Genetic Diversity among Strains of Mycoplasma hyopneumoniae
    Article Snippet: These assays were then used to test DNA from each of the isolates in Table . .. Some assays were performed as previously published ( , , ), while others ( , , , , ) were modified by substituting MasterAmp E buffer (Epicentre Biotechnologies, Madison, WI) for the published buffer with their published cycling parameters in reaction mixtures consisting of 1× Master Amp E, 0.4 mM each forward and reverse primer, 2.5 U of Taq (New England Biolabs, Inc., Ipswich, ME), template containing 1 to 10 ng of DNA, and a sufficient quantity of PCR-grade water to a final volume of 25 μl. .. The primer pairs from nested-set PCRs were run as individual assays (not as nested sets) to independently evaluate the ability of each primer pair to form products.

    Article Title: Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments
    Article Snippet: When the plants reached the 1 to 2 leaf stage, 3 cm leaf segments from primary leaves were harvested for genomic DNA isolation. .. A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process. .. The PCR conditions were: initial denaturation at 94 °C (3 min), followed by 44 cycles of 94 °C (1 min), 55 or 60 °C annealing (1 min), and 72 °C extension (1 min) with a final extension at 72 °C for 10 min.

    Article Title: Ocean currents influence the genetic structure of an intertidal mollusc in southeastern Australia - implications for predicting the movement of passive dispersers across a marine biogeographic barrier
    Article Snippet: Approximately 650 base pairs (bp) of the mitochondrial cytochrome oxidase I gene (COI ) was amplified for 10 individuals from each sample site by PCR using primers HCO and LCO (Folmer et al. ). .. PCRs were prepared in 25 μL volumes each containing 10.25 μL ddH2O, 1× reaction buffer, 5.0 μL dNTPs (1 mmol/L), 0.4 μmol/L HCO primer (10 μmol/L), 0.4 μmol/L LCO primer (10 μmol/L), 0.25 units NEB taq (Biosciences, New England), and 5 μL DNA extract.

    Article Title: CRISPR-MVLST subtyping of Salmonella enterica subsp. enterica serovars Typhimurium and Heidelberg and application in identifying outbreak isolates
    Article Snippet: Primers for amplification of all four genomic loci are listed in Table . .. PCR reactions were performed in a total volume of 25 μl: 1.5 μl template, 0.3 μl Taq (1.5 units; New England Bio Labs, Ipswich, MA), 0.2 μl 10 mM dNTPs, 1 μl of each 10 μM primer, 2.5 μl of 10× Taq buffer and 18.5 μl water. .. PCR conditions were as follows and the annealing temperatures (AT) are listed in Table : initial denaturation step of 10 minutes at 94°C followed by 35 cycles of 1 minute at 94°C, 1 minute at AT and extension for 1 minute (fimH and sseL ) or 1.5 minutes (CRISPR1 and CRISPR2) at 72°C; a final extension step was done at 72°C for 8 minutes.

    Article Title: RAD marker microarrays enable rapid mapping of zebrafish mutations
    Article Snippet: Genomic DNA samples were digested with Eco RI (New England Biolabs, Ipswich, MA, USA) and short linkers (5'-GACTCCTCGACTCTCACATCTGGACATA-3', 5'-Phos-AATTTATGTCCAGATGTGAGAGTC-3') were ligated to the fragment ends using previously described conditions [ ]. .. PCR reactions were carried out in 50 μl under the following conditions: 50 ng template, 1 × Thermopol Buffer (New England Biolabs), 2.5 U Taq (New England Biolabs), 0.2 mmol/l dNTPs (Fermentas, Burlington, Ontario, Canada), 0.4 μmol/l forward primer, and 0.2 μmol/l reverse primer. .. Touchdown amplification was applied, with primer annealing temperature dropping 2°C every other cycle from 65°C to 55°C, after which 25 additional cycles were carried out [ ].

    Article Title: A Simple Isothermal DNA Amplification Method to Screen Black Flies for Onchocerca volvulus Infection
    Article Snippet: For determination of amplification measured by color change (purple to sky blue), 0.15 µL of 120 µM hydroxy naphthol blue (HNB, Sigma-Aldrich Inc, St. Louis, MO, USA) was added to the reaction mixture. .. LAMP primers B3 and F3 were used to PCR amplify OvGST1a in 25 µL reactions containing 3 µL DNA template, 0.2 µM of each primer, and 1.25 U of Taq DNA polymerase in 1× standard buffer (New England Biolabs) containing 3.5 mM MgCl2 , 0.2 mM and 0.2 mM dNTP each. .. All reactions were denatured once at 94°C for 5 min followed by 35 cycles of the following cycling conditions: 30 s at 94°C, 1 min at 53°C, 1 min at 72°C, and a final extension for 5 min at 72°C using a Gene Amp PCR system 9700 (Applied Biosystems).

    Article Title: Cooperative activity of GABP with PU.1 or C/EBPε regulates lamin B receptor gene expression, implicating their roles in granulocyte nuclear maturation
    Article Snippet: Data analysis was performed using the ΔΔCT method. .. A 25 μl PCR reaction was also performed using 2 μl of purified DNA from EML cells using 1X standard Taq reaction buffer (New England BioLabs), 1 U of Taq DNA polymerase, plus 0.2 pmol/μl of same forward and reverse primers for proximal and distal Ets sites as mentioned above. .. PCR was carried out for 28 cycles by using 1 cycle of 95°C for 1 min; 30 cycles of 95°C, 63°C and 72°C each at 30 s and finally 1 cycle of 72°C for 5 min. Five microliters of PCR product was electrophoresed on 0.9 % agarose gel and bands were visualized by staining the gel in ethidium bromide.

    Article Title: 16S rRNA Terminal Restriction Fragment Length Polymorphism for the Characterization of the Nasopharyngeal Microbiota
    Article Snippet: Purified PCR products were checked for correct amplification size and purity using the Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, CA) with the DNA 7500 LabChip kit. .. The enzymatic digestions were performed in a total volume of 20 µl using 40 ng of the amplified and purified PCR product, 1 µl NEB Reaction Buffer (New England Biolabs, Ipswich, MA) 1 (for digestion with Tsp509I ) or 4 (Hpy166II ) and 0.5 µl of the restriction enzymes Tsp509I or Hpy166II . .. Digestion conditions were 3 h at 65°C for TSP509I and 3 h at 37°C with 20 min enzyme inactivation at 65°C for Hpy166II , respectively.

    Article Title: A new insect cell glycoengineering approach provides baculovirus-inducible glycogene expression and increases human-type glycosylation efficiency
    Article Snippet: The RNA preparations were treated with RNAse-free DNAse I (New England Biolabs), and then 400 ng of each were reverse transcribed at 50°C for 30 min with Thermoscript Reverse Transcriptase (Life Technologies) and oligo(dT)31 -VN (5′-TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTVN-3′) as the primer. .. The resulting cDNA preparations were treated with RNAse H and used for PCR reactions with Taq Polymerase and Crimson Taq Buffer (New England Biolabs). .. The PCR conditions included an initial denaturation step of 95°C for 30 seconds, followed by 30 (MGAT1, MGAT2, B4GALT1, SAS, ST3GAL4b, SfRPL3) or 33 (GNPE, CMAS, CSAT, ST6GAL1) cycles of denaturation at 95°C for 15 seconds, annealing at 50°C for 20 seconds, and extension at 68°C for 30 seconds.

    Peptide Microarray:

    Article Title: Protein-Tyrosine Kinase Activity Profiling in Knock Down Zebrafish Embryos
    Article Snippet: Two N-terminal residues link the phosphosite sequence to the solid support of the 3-dimensional chip. .. The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array. .. Prior to application of the sample, the chips were blocked using a solution of 2% BSA (Bovine Serum Albumin, Fraction V, Calbiochem, Germany), and washed 2 times using kinase reaction buffer.

    Imaging:

    Article Title: Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments
    Article Snippet: A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process. .. The amplification products were resolved by capillary electrophoresis using an ABI3130XL DNA fragment analyser (Applied Biosystems), or by horizontal high resolution agarose electrophoresis using 2 % Metaphor and 1 % agarose LE gels at 4 V cm−1 in TBE (0.045 M TRIS, 0.045 M Borate, and 0.001 M EDTA) buffer and stained with ethidium bromide (0.5 μg ml−1 ).

    Article Title: Protein-Tyrosine Kinase Activity Profiling in Knock Down Zebrafish Embryos
    Article Snippet: Data was captured by real-time imaging of the fluorescence signal by CCD imaging , . .. The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array.

    Sequencing:

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: Paragraph title: PCR-GBS library preparation and ion torrent sequencing ... The barcoding PCR reaction consisted of 0.235 µl of H2 O, 0.625 µl NEB Standard Taq Buffer (1.25 ×), 0.1 µl dNTPs (500 µM), 1 µl universal reverse primer (1 µM), 1 µl unique barcode adaptor primer (0.4 µM), 0.04 µl of NEB Taq polymerase (0.2 units), and 2 µl template (diluted primary PCR reaction).

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions. .. The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions.

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: Paragraph title: Preparation of sequencing libraries ... The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr.

    Article Title: Impact of swapping soils on the endophytic bacterial communities of pre-domesticated, ancient and modern maize
    Article Snippet: Total DNA from 250 mg of soil or sand was extracted using Powersoil DNA isolation kits (MoBio, USA), and DNA was quantified using a Nanodrop (Thermo Scientific, USA). .. A PCR mastermix was made with the following components per 25 μl volume: 2.5 μl Standard Taq Buffer (New England Biolabs), 0.5 μl of 25 mM dNTP mix, 0.5 μl of 10 mM 27 F-Degen primer with sequence AGRRTTYGATYMTGGYTYAG [ ] (where R = A + G, Y = C + T, M = A + C), 0.5 μl of 10 mM 1492r primer with sequence GGTTACCTTGTTACGACTT [ ], 0.25 μl of 50 mM MgCl2 , 0.25 μl of Standard Taq (New England Biolabs), 50 ng of total DNA, and double distilled water up to 25 μl total. .. Amplification was for 35 cycles in a PTC200 DNA Thermal Cycler (MJ Scientific, USA) using the following program: 96°C for 3 min, 35X (94°C for 30 sec, 48°C for 30 sec, 72°C for 1:30 min), 72°C for 7 min.

    Article Title: A Recurrent Stop-Codon Mutation in Succinate Dehydrogenase Subunit B Gene in Normal Peripheral Blood and Childhood T-Cell Acute Leukemia
    Article Snippet: Fifteen µL of the fluorescently-labeled PCR products, which contained ∼200–300 ng of DNA, were directly digested by 10 U of Taq I RE, 1× Taq I RE buffer and 1× Bovine Serum Albumin (all from New England Biolabs) overnight in a 25 µL reaction volume. .. The fluorescently-labeled and Taq I-RE digested PCR products were run in capillary gel electrophoresis (3100 Avant Genetic Analyzer, ABI Prism, Applied Biosystems/HITACHI).

    Article Title: Allopatric Speciation within a Cryptic Species Complex of Australasian Octopuses
    Article Snippet: Paragraph title: Sequencing ... 25 µL reactions comprised 0.1 µL Taq (Onetaq, New England Biolabs ), 2.5 µL 10 x buffer (Paq5000™), 2 µL dNTP mix (10 µM, Bioline ), 0.5 µL forward primer (10 µM), 0.5 µL reverse primer (10 µM), 17.4 µL ddH2 O and 2 µL DNA (diluted to between 1–5 ng/µL).

    Article Title: RAD marker microarrays enable rapid mapping of zebrafish mutations
    Article Snippet: PCR reactions were carried out in 50 μl under the following conditions: 50 ng template, 1 × Thermopol Buffer (New England Biolabs), 2.5 U Taq (New England Biolabs), 0.2 mmol/l dNTPs (Fermentas, Burlington, Ontario, Canada), 0.4 μmol/l forward primer, and 0.2 μmol/l reverse primer. .. PCR reactions were carried out in 50 μl under the following conditions: 50 ng template, 1 × Thermopol Buffer (New England Biolabs), 2.5 U Taq (New England Biolabs), 0.2 mmol/l dNTPs (Fermentas, Burlington, Ontario, Canada), 0.4 μmol/l forward primer, and 0.2 μmol/l reverse primer.

    Article Title: Protein-Tyrosine Kinase Activity Profiling in Knock Down Zebrafish Embryos
    Article Snippet: Two N-terminal residues link the phosphosite sequence to the solid support of the 3-dimensional chip. .. The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array.

    Staining:

    Article Title: Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments
    Article Snippet: A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process. .. A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process.

    Article Title: A Simple Isothermal DNA Amplification Method to Screen Black Flies for Onchocerca volvulus Infection
    Article Snippet: LAMP primers B3 and F3 were used to PCR amplify OvGST1a in 25 µL reactions containing 3 µL DNA template, 0.2 µM of each primer, and 1.25 U of Taq DNA polymerase in 1× standard buffer (New England Biolabs) containing 3.5 mM MgCl2 , 0.2 mM and 0.2 mM dNTP each. .. All reactions were denatured once at 94°C for 5 min followed by 35 cycles of the following cycling conditions: 30 s at 94°C, 1 min at 53°C, 1 min at 72°C, and a final extension for 5 min at 72°C using a Gene Amp PCR system 9700 (Applied Biosystems).

    Article Title: Vitamin E inhibits the UVAI induction of “light” and “dark” cyclobutane pyrimidine dimers, and oxidatively generated DNA damage, in keratinocytes
    Article Snippet: T4endoV (0.1 U/mL), or hOGG1 (3.2 U/mL), or enzyme reaction buffer alone, was added to each gel (both enzymes were purchased from New England Biolabs, Hitchin, UK). .. Finally, slides were rinsed with neutralization solution (0.4 M Trizma Base, pH 7.5; Sigma) for 20 min and then washed with ddH2 O for 10 min.

    DNA Extraction:

    Article Title: Impact of swapping soils on the endophytic bacterial communities of pre-domesticated, ancient and modern maize
    Article Snippet: Paragraph title: DNA extraction and Terminal Restriction Fragment Length Polymorphism (TRFLP) from plant tissues and soil ... A PCR mastermix was made with the following components per 25 μl volume: 2.5 μl Standard Taq Buffer (New England Biolabs), 0.5 μl of 25 mM dNTP mix, 0.5 μl of 10 mM 27 F-Degen primer with sequence AGRRTTYGATYMTGGYTYAG [ ] (where R = A + G, Y = C + T, M = A + C), 0.5 μl of 10 mM 1492r primer with sequence GGTTACCTTGTTACGACTT [ ], 0.25 μl of 50 mM MgCl2 , 0.25 μl of Standard Taq (New England Biolabs), 50 ng of total DNA, and double distilled water up to 25 μl total.

    Article Title: Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments
    Article Snippet: When the plants reached the 1 to 2 leaf stage, 3 cm leaf segments from primary leaves were harvested for genomic DNA isolation. .. A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process.

    Nucleic Acid Electrophoresis:

    Article Title: A Recurrent Stop-Codon Mutation in Succinate Dehydrogenase Subunit B Gene in Normal Peripheral Blood and Childhood T-Cell Acute Leukemia
    Article Snippet: Paragraph title: Quantification of the R46X mutation by TaqI RE digestion and capillary gel electrophoresis ... Fifteen µL of the fluorescently-labeled PCR products, which contained ∼200–300 ng of DNA, were directly digested by 10 U of Taq I RE, 1× Taq I RE buffer and 1× Bovine Serum Albumin (all from New England Biolabs) overnight in a 25 µL reaction volume.

    Fluorescence:

    Article Title: Protein-Tyrosine Kinase Activity Profiling in Knock Down Zebrafish Embryos
    Article Snippet: Data was captured by real-time imaging of the fluorescence signal by CCD imaging , . .. The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array.

    Mutagenesis:

    Article Title: Prp8 retinitis pigmentosa mutants cause defects in the transition between the catalytic steps of splicing
    Article Snippet: For qPCR, 10 µL of diluted cDNA from reverse transcription was combined with 0.25 mM each gene-specific primer, 1× SYBR green (Roche), 0.625 µL dimethyl sulfoxide (Invitrogen), 0.4 mM dTNPs (Fermentas), 1× Standard Taq Buffer (NEB) and 0.25 µL Standard Taq (NEB) and water to a total volume of 25 µL. qPCR cycle conditions were 95°C 3 min, 39 cycles of 95°C 15 sec, 55°C 30 sec, 72°C 15 sec, followed by one cycle at 72°C for 3 min. Exon primers were designed to be within the 3′ exon of all genes tested and were used to measure total RNA levels. pre-mRNA levels were assessed using intron primers. .. For qPCR, 10 µL of diluted cDNA from reverse transcription was combined with 0.25 mM each gene-specific primer, 1× SYBR green (Roche), 0.625 µL dimethyl sulfoxide (Invitrogen), 0.4 mM dTNPs (Fermentas), 1× Standard Taq Buffer (NEB) and 0.25 µL Standard Taq (NEB) and water to a total volume of 25 µL. qPCR cycle conditions were 95°C 3 min, 39 cycles of 95°C 15 sec, 55°C 30 sec, 72°C 15 sec, followed by one cycle at 72°C for 3 min. Exon primers were designed to be within the 3′ exon of all genes tested and were used to measure total RNA levels. pre-mRNA levels were assessed using intron primers.

    Article Title: A Recurrent Stop-Codon Mutation in Succinate Dehydrogenase Subunit B Gene in Normal Peripheral Blood and Childhood T-Cell Acute Leukemia
    Article Snippet: Paragraph title: Quantification of the R46X mutation by TaqI RE digestion and capillary gel electrophoresis ... Fifteen µL of the fluorescently-labeled PCR products, which contained ∼200–300 ng of DNA, were directly digested by 10 U of Taq I RE, 1× Taq I RE buffer and 1× Bovine Serum Albumin (all from New England Biolabs) overnight in a 25 µL reaction volume.

    Isolation:

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: Polyadenylated RNA was purified from 10 µg of total RNA using the Magnetic mRNA Isolation Kit (New England Biolabs, MA). .. The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr.

    Article Title: Prp8 retinitis pigmentosa mutants cause defects in the transition between the catalytic steps of splicing
    Article Snippet: Paragraph title: RNA isolation and RT-qPCR ... For qPCR, 10 µL of diluted cDNA from reverse transcription was combined with 0.25 mM each gene-specific primer, 1× SYBR green (Roche), 0.625 µL dimethyl sulfoxide (Invitrogen), 0.4 mM dTNPs (Fermentas), 1× Standard Taq Buffer (NEB) and 0.25 µL Standard Taq (NEB) and water to a total volume of 25 µL. qPCR cycle conditions were 95°C 3 min, 39 cycles of 95°C 15 sec, 55°C 30 sec, 72°C 15 sec, followed by one cycle at 72°C for 3 min. Exon primers were designed to be within the 3′ exon of all genes tested and were used to measure total RNA levels. pre-mRNA levels were assessed using intron primers.

    Size-exclusion Chromatography:

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: The primary PCR reaction was conducted as follows: Initial denaturation at 94° for 10 min, 10 cycles of 94° for 20 sec and 64° decreasing each cycle by 0.8° for 60 sec, followed by 20 cycles of 94° for 20 sec, 57° for 60 sec, and 68° for 30 sec, ending with a final extension of 72° for 3 min. .. The barcoding PCR reaction consisted of 0.235 µl of H2 O, 0.625 µl NEB Standard Taq Buffer (1.25 ×), 0.1 µl dNTPs (500 µM), 1 µl universal reverse primer (1 µM), 1 µl unique barcode adaptor primer (0.4 µM), 0.04 µl of NEB Taq polymerase (0.2 units), and 2 µl template (diluted primary PCR reaction).

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: The parameters for the PCR reactions were as follows: 94° for 5 min, followed by 35 cycles of 94° for 30 sec, 62° for 30 sec, and 72° for 60 sec, followed by a final extension at 72° for 7 min. .. The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions.

    Article Title: Prp8 retinitis pigmentosa mutants cause defects in the transition between the catalytic steps of splicing
    Article Snippet: Reactions were then diluted between 1:75 and 1:250 with water prior to analysis by qPCR. .. For qPCR, 10 µL of diluted cDNA from reverse transcription was combined with 0.25 mM each gene-specific primer, 1× SYBR green (Roche), 0.625 µL dimethyl sulfoxide (Invitrogen), 0.4 mM dTNPs (Fermentas), 1× Standard Taq Buffer (NEB) and 0.25 µL Standard Taq (NEB) and water to a total volume of 25 µL. qPCR cycle conditions were 95°C 3 min, 39 cycles of 95°C 15 sec, 55°C 30 sec, 72°C 15 sec, followed by one cycle at 72°C for 3 min. Exon primers were designed to be within the 3′ exon of all genes tested and were used to measure total RNA levels. pre-mRNA levels were assessed using intron primers. .. The product of these primes spanned an intron–exon junction to ensure that only pre-mRNA was measured.

    Purification:

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: The barcoding PCR reaction consisted of 0.235 µl of H2 O, 0.625 µl NEB Standard Taq Buffer (1.25 ×), 0.1 µl dNTPs (500 µM), 1 µl universal reverse primer (1 µM), 1 µl unique barcode adaptor primer (0.4 µM), 0.04 µl of NEB Taq polymerase (0.2 units), and 2 µl template (diluted primary PCR reaction). .. Following PCR, the plate was briefly centrifuged and each sample was diluted by adding 15 µl of H2 O.

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: Fragmented cDNA was purified and the ends were repaired using NEB Quick Blunting Kit (New England Biolabs, MA) according to manufacturer’s protocol. .. The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr. .. Tailed templates were ligated to double stranded adaptors prepared with oligonucleotides from the Illumina Customer Sequence Letter (version August 12, 2014, ; Table S1 ).

    Article Title: Cooperative activity of GABP with PU.1 or C/EBPε regulates lamin B receptor gene expression, implicating their roles in granulocyte nuclear maturation
    Article Snippet: Data analysis was performed using the ΔΔCT method. .. A 25 μl PCR reaction was also performed using 2 μl of purified DNA from EML cells using 1X standard Taq reaction buffer (New England BioLabs), 1 U of Taq DNA polymerase, plus 0.2 pmol/μl of same forward and reverse primers for proximal and distal Ets sites as mentioned above. .. PCR was carried out for 28 cycles by using 1 cycle of 95°C for 1 min; 30 cycles of 95°C, 63°C and 72°C each at 30 s and finally 1 cycle of 72°C for 5 min. Five microliters of PCR product was electrophoresed on 0.9 % agarose gel and bands were visualized by staining the gel in ethidium bromide.

    Article Title: 16S rRNA Terminal Restriction Fragment Length Polymorphism for the Characterization of the Nasopharyngeal Microbiota
    Article Snippet: Purified PCR products were checked for correct amplification size and purity using the Agilent 2100 bioanalyzer (Agilent Technologies, Palo Alto, CA) with the DNA 7500 LabChip kit. .. The enzymatic digestions were performed in a total volume of 20 µl using 40 ng of the amplified and purified PCR product, 1 µl NEB Reaction Buffer (New England Biolabs, Ipswich, MA) 1 (for digestion with Tsp509I ) or 4 (Hpy166II ) and 0.5 µl of the restriction enzymes Tsp509I or Hpy166II . .. Digestion conditions were 3 h at 65°C for TSP509I and 3 h at 37°C with 20 min enzyme inactivation at 65°C for Hpy166II , respectively.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: A new insect cell glycoengineering approach provides baculovirus-inducible glycogene expression and increases human-type glycosylation efficiency
    Article Snippet: Paragraph title: 2.5. RT-PCR assays ... The resulting cDNA preparations were treated with RNAse H and used for PCR reactions with Taq Polymerase and Crimson Taq Buffer (New England Biolabs).

    Quantitative RT-PCR:

    Article Title: Prp8 retinitis pigmentosa mutants cause defects in the transition between the catalytic steps of splicing
    Article Snippet: Paragraph title: RNA isolation and RT-qPCR ... For qPCR, 10 µL of diluted cDNA from reverse transcription was combined with 0.25 mM each gene-specific primer, 1× SYBR green (Roche), 0.625 µL dimethyl sulfoxide (Invitrogen), 0.4 mM dTNPs (Fermentas), 1× Standard Taq Buffer (NEB) and 0.25 µL Standard Taq (NEB) and water to a total volume of 25 µL. qPCR cycle conditions were 95°C 3 min, 39 cycles of 95°C 15 sec, 55°C 30 sec, 72°C 15 sec, followed by one cycle at 72°C for 3 min. Exon primers were designed to be within the 3′ exon of all genes tested and were used to measure total RNA levels. pre-mRNA levels were assessed using intron primers.

    Lysis:

    Article Title: Vitamin E inhibits the UVAI induction of “light” and “dark” cyclobutane pyrimidine dimers, and oxidatively generated DNA damage, in keratinocytes
    Article Snippet: After removing the lysis buffer, slides were washed with ice-cold ddH2 O for 10 min (in the dark, to prevent adventitious DNA damage). .. T4endoV (0.1 U/mL), or hOGG1 (3.2 U/mL), or enzyme reaction buffer alone, was added to each gel (both enzymes were purchased from New England Biolabs, Hitchin, UK).

    Nested PCR:

    Article Title: Polymorphisms in K13 and Falcipain-2 Associated with Artemisinin Resistance Are Not Prevalent in Plasmodium falciparum Isolated from Ugandan Children
    Article Snippet: For the FP2 gene, 2 nested PCR reactions were utilized to amplify the entire gene, spanning nucleotides 1–873 and 271–1455. .. For both loci, first and second round amplifications were performed in 25 µl reactions containing 160 nM primers (Integrated DNA Technologies), 160 µM dNTPs (Invitrogen), 1 unit Taq DNA Polymerase (New England BioLabs), and 2 µl DNA/primary reaction template in 1x Standard Taq Buffer (New England BioLabs).

    Article Title: Impact of swapping soils on the endophytic bacterial communities of pre-domesticated, ancient and modern maize
    Article Snippet: A PCR mastermix was made with the following components per 25 μl volume: 2.5 μl Standard Taq Buffer (New England Biolabs), 0.5 μl of 25 mM dNTP mix, 0.5 μl of 10 mM 27 F-Degen primer with sequence AGRRTTYGATYMTGGYTYAG [ ] (where R = A + G, Y = C + T, M = A + C), 0.5 μl of 10 mM 1492r primer with sequence GGTTACCTTGTTACGACTT [ ], 0.25 μl of 50 mM MgCl2 , 0.25 μl of Standard Taq (New England Biolabs), 50 ng of total DNA, and double distilled water up to 25 μl total. .. A PCR mastermix was made with the following components per 25 μl volume: 2.5 μl Standard Taq Buffer (New England Biolabs), 0.5 μl of 25 mM dNTP mix, 0.5 μl of 10 mM 27 F-Degen primer with sequence AGRRTTYGATYMTGGYTYAG [ ] (where R = A + G, Y = C + T, M = A + C), 0.5 μl of 10 mM 1492r primer with sequence GGTTACCTTGTTACGACTT [ ], 0.25 μl of 50 mM MgCl2 , 0.25 μl of Standard Taq (New England Biolabs), 50 ng of total DNA, and double distilled water up to 25 μl total.

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments
    Article Snippet: When the plants reached the 1 to 2 leaf stage, 3 cm leaf segments from primary leaves were harvested for genomic DNA isolation. .. A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process. .. The PCR conditions were: initial denaturation at 94 °C (3 min), followed by 44 cycles of 94 °C (1 min), 55 or 60 °C annealing (1 min), and 72 °C extension (1 min) with a final extension at 72 °C for 10 min.

    Chromatin Immunoprecipitation:

    Article Title: Cooperative activity of GABP with PU.1 or C/EBPε regulates lamin B receptor gene expression, implicating their roles in granulocyte nuclear maturation
    Article Snippet: Paragraph title: Chromatin Immunoprecipitation (ChIP) and real-time PCR assay ... A 25 μl PCR reaction was also performed using 2 μl of purified DNA from EML cells using 1X standard Taq reaction buffer (New England BioLabs), 1 U of Taq DNA polymerase, plus 0.2 pmol/μl of same forward and reverse primers for proximal and distal Ets sites as mentioned above.

    Article Title: Protein-Tyrosine Kinase Activity Profiling in Knock Down Zebrafish Embryos
    Article Snippet: Two N-terminal residues link the phosphosite sequence to the solid support of the 3-dimensional chip. .. The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array.

    Plasmid Preparation:

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions. .. The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions.

    Software:

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions. .. The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions.

    Article Title: Protein-Tyrosine Kinase Activity Profiling in Knock Down Zebrafish Embryos
    Article Snippet: The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array. .. The zebrafish embryo lysates were analysed by applying an aliquot of the lysate in kinase reaction buffer (Abl reaction buffer [New England Biolabs], consisting of 100 mM MgCl2 , 10 mM EGTA, 20 mM DTT and 0.1% Brij 35 in 500 mM Tris/HCl, pH 7.5) containing 12.5 µg/ml fluoresceine labelled PY20 antibody against phosphotyrosine (Exalpha, USA), and 400 µM ATP (Sigma) to the PamChip peptide array.

    Real-time Polymerase Chain Reaction:

    Article Title: Prp8 retinitis pigmentosa mutants cause defects in the transition between the catalytic steps of splicing
    Article Snippet: Reactions were then diluted between 1:75 and 1:250 with water prior to analysis by qPCR. .. For qPCR, 10 µL of diluted cDNA from reverse transcription was combined with 0.25 mM each gene-specific primer, 1× SYBR green (Roche), 0.625 µL dimethyl sulfoxide (Invitrogen), 0.4 mM dTNPs (Fermentas), 1× Standard Taq Buffer (NEB) and 0.25 µL Standard Taq (NEB) and water to a total volume of 25 µL. qPCR cycle conditions were 95°C 3 min, 39 cycles of 95°C 15 sec, 55°C 30 sec, 72°C 15 sec, followed by one cycle at 72°C for 3 min. Exon primers were designed to be within the 3′ exon of all genes tested and were used to measure total RNA levels. pre-mRNA levels were assessed using intron primers. .. The product of these primes spanned an intron–exon junction to ensure that only pre-mRNA was measured.

    Article Title: Cooperative activity of GABP with PU.1 or C/EBPε regulates lamin B receptor gene expression, implicating their roles in granulocyte nuclear maturation
    Article Snippet: Paragraph title: Chromatin Immunoprecipitation (ChIP) and real-time PCR assay ... A 25 μl PCR reaction was also performed using 2 μl of purified DNA from EML cells using 1X standard Taq reaction buffer (New England BioLabs), 1 U of Taq DNA polymerase, plus 0.2 pmol/μl of same forward and reverse primers for proximal and distal Ets sites as mentioned above.

    Multiplex Assay:

    Article Title: De Novo Assembly and Characterization of Four Anthozoan (Phylum Cnidaria) Transcriptomes
    Article Snippet: The product was purified and A-tailed in a reaction with nuclease-free water, 1× NEB Standard Taq buffer (New England Biolabs, MA), 1 mM dATP, and 2 units of NEB Standard Taq (New England Biolabs, MA) at 68° for 2 hr. .. Ligation products were purified and then amplified using custom sample-specific barcodes (“indices”) designed with a 3-bp minimum Hamming distance based on Illumina barcodes ( ) ( Table S1 ).

    Selection:

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: A set of these ESTs were selected at ∼60 kb intervals using predicted function as the criteria for selection, as certain genes have a higher probability of containing polymorphisms compared to others. .. The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions.

    Agarose Gel Electrophoresis:

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: Additionally, 2 µL of each reaction was separated on a 1% agarose gel containing GelRed to ensure successful reactions. .. The barcoding PCR reaction consisted of 0.235 µl of H2 O, 0.625 µl NEB Standard Taq Buffer (1.25 ×), 0.1 µl dNTPs (500 µM), 1 µl universal reverse primer (1 µM), 1 µl unique barcode adaptor primer (0.4 µM), 0.04 µl of NEB Taq polymerase (0.2 units), and 2 µl template (diluted primary PCR reaction).

    Article Title: CRISPR-MVLST subtyping of Salmonella enterica subsp. enterica serovars Typhimurium and Heidelberg and application in identifying outbreak isolates
    Article Snippet: PCR reactions were performed in a total volume of 25 μl: 1.5 μl template, 0.3 μl Taq (1.5 units; New England Bio Labs, Ipswich, MA), 0.2 μl 10 mM dNTPs, 1 μl of each 10 μM primer, 2.5 μl of 10× Taq buffer and 18.5 μl water. .. PCR conditions were as follows and the annealing temperatures (AT) are listed in Table : initial denaturation step of 10 minutes at 94°C followed by 35 cycles of 1 minute at 94°C, 1 minute at AT and extension for 1 minute (fimH and sseL ) or 1.5 minutes (CRISPR1 and CRISPR2) at 72°C; a final extension step was done at 72°C for 8 minutes.

    Article Title: RAD marker microarrays enable rapid mapping of zebrafish mutations
    Article Snippet: PCR reactions were carried out in 50 μl under the following conditions: 50 ng template, 1 × Thermopol Buffer (New England Biolabs), 2.5 U Taq (New England Biolabs), 0.2 mmol/l dNTPs (Fermentas, Burlington, Ontario, Canada), 0.4 μmol/l forward primer, and 0.2 μmol/l reverse primer. .. Touchdown amplification was applied, with primer annealing temperature dropping 2°C every other cycle from 65°C to 55°C, after which 25 additional cycles were carried out [ ].

    Article Title: A Simple Isothermal DNA Amplification Method to Screen Black Flies for Onchocerca volvulus Infection
    Article Snippet: LAMP primers B3 and F3 were used to PCR amplify OvGST1a in 25 µL reactions containing 3 µL DNA template, 0.2 µM of each primer, and 1.25 U of Taq DNA polymerase in 1× standard buffer (New England Biolabs) containing 3.5 mM MgCl2 , 0.2 mM and 0.2 mM dNTP each. .. All reactions were denatured once at 94°C for 5 min followed by 35 cycles of the following cycling conditions: 30 s at 94°C, 1 min at 53°C, 1 min at 72°C, and a final extension for 5 min at 72°C using a Gene Amp PCR system 9700 (Applied Biosystems).

    Electrophoresis:

    Article Title: Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments
    Article Snippet: A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process. .. A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process.

    Article Title: A Simple Isothermal DNA Amplification Method to Screen Black Flies for Onchocerca volvulus Infection
    Article Snippet: LAMP primers B3 and F3 were used to PCR amplify OvGST1a in 25 µL reactions containing 3 µL DNA template, 0.2 µM of each primer, and 1.25 U of Taq DNA polymerase in 1× standard buffer (New England Biolabs) containing 3.5 mM MgCl2 , 0.2 mM and 0.2 mM dNTP each. .. All reactions were denatured once at 94°C for 5 min followed by 35 cycles of the following cycling conditions: 30 s at 94°C, 1 min at 53°C, 1 min at 72°C, and a final extension for 5 min at 72°C using a Gene Amp PCR system 9700 (Applied Biosystems).

    Article Title: 16S rRNA Terminal Restriction Fragment Length Polymorphism for the Characterization of the Nasopharyngeal Microbiota
    Article Snippet: The enzymatic digestions were performed in a total volume of 20 µl using 40 ng of the amplified and purified PCR product, 1 µl NEB Reaction Buffer (New England Biolabs, Ipswich, MA) 1 (for digestion with Tsp509I ) or 4 (Hpy166II ) and 0.5 µl of the restriction enzymes Tsp509I or Hpy166II . .. The enzymatic digestions were performed in a total volume of 20 µl using 40 ng of the amplified and purified PCR product, 1 µl NEB Reaction Buffer (New England Biolabs, Ipswich, MA) 1 (for digestion with Tsp509I ) or 4 (Hpy166II ) and 0.5 µl of the restriction enzymes Tsp509I or Hpy166II .

    Article Title: Vitamin E inhibits the UVAI induction of “light” and “dark” cyclobutane pyrimidine dimers, and oxidatively generated DNA damage, in keratinocytes
    Article Snippet: T4endoV (0.1 U/mL), or hOGG1 (3.2 U/mL), or enzyme reaction buffer alone, was added to each gel (both enzymes were purchased from New England Biolabs, Hitchin, UK). .. T4endoV (0.1 U/mL), or hOGG1 (3.2 U/mL), or enzyme reaction buffer alone, was added to each gel (both enzymes were purchased from New England Biolabs, Hitchin, UK).

    Concentration Assay:

    Article Title: Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments
    Article Snippet: When the plants reached the 1 to 2 leaf stage, 3 cm leaf segments from primary leaves were harvested for genomic DNA isolation. .. A 10 μl PCR reaction consisting of DNA (final concentration of 2.4 ng/μl), Ultrapure double distilled H2 0 (Gibco), 10 % PCR Buffer without MgCl2 [Invitrogen cat.#18067-017: 200 mM Tris–HCl (pH 8.4), 500 mM KCl], 10 mM dNTPs (Roche), 1.5 mM MgCl2 (Invitrogen), 0.07 U μl−1 Taq (5 U of activity μl−1 ) NEB, and 2 ηg μl−1 forward and 2 ηg μl−1 reverse primer was used for the DNA amplification process. .. The PCR conditions were: initial denaturation at 94 °C (3 min), followed by 44 cycles of 94 °C (1 min), 55 or 60 °C annealing (1 min), and 72 °C extension (1 min) with a final extension at 72 °C for 10 min.

    Article Title: Cooperative activity of GABP with PU.1 or C/EBPε regulates lamin B receptor gene expression, implicating their roles in granulocyte nuclear maturation
    Article Snippet: Real-time PCR was performed by using 2 μl of purified DNA as template, SsoAdvanced SYBR Green Supermix (Bio-Rad) and the following primers at a final concentration of 0.2 pmol/μl: Proximal Ets site: (F) 5′-AACAGCACGGCGAAGTAG-3′ and (R) 5′-CCCCTTACACTTGCCACC-3′, Distal Ets site: (F) 5′-GATCTGTTTGTCCCCAGCTAC-3′ and (R) 5′-ACTTTGTGAGCACTTGAGACC-3′, and Proximal Cebp site: (F) 5′-CTCGGAGTAGGATTCGTCTTTAAG-3′ and (R) 5′-GATTTGTCATTGCCGTTGGG-3′. .. A 25 μl PCR reaction was also performed using 2 μl of purified DNA from EML cells using 1X standard Taq reaction buffer (New England BioLabs), 1 U of Taq DNA polymerase, plus 0.2 pmol/μl of same forward and reverse primers for proximal and distal Ets sites as mentioned above.

    Marker:

    Article Title: Fine Mapping of the Barley Chromosome 6H Net Form Net Blotch Susceptibility Locus
    Article Snippet: Paragraph title: STS/SNP marker development ... The PCR reactions consisted of 1.25 units NEB Standard Taq polymerase, forward and reverse primers (1.2 µM), NEB Standard Taq buffer (1 ×), and dNTPs (200 µM), in 25 µl reactions.

    Single Cell Gel Electrophoresis:

    Article Title: Vitamin E inhibits the UVAI induction of “light” and “dark” cyclobutane pyrimidine dimers, and oxidatively generated DNA damage, in keratinocytes
    Article Snippet: Paragraph title: Measurement of DNA damage by the comet assay ... T4endoV (0.1 U/mL), or hOGG1 (3.2 U/mL), or enzyme reaction buffer alone, was added to each gel (both enzymes were purchased from New England Biolabs, Hitchin, UK).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs standard taq reaction buffer
    Standard Taq Reaction Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/standard taq reaction buffer/product/New England Biolabs
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    standard taq reaction buffer - by Bioz Stars, 2019-11
    99/100 stars
      Buy from Supplier

    Image Search Results