taqman genotyping master mix  (New England Biolabs)


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    Name:
    OneTaq Hot Start 2X Master Mix with Standard Buffer
    Description:
    OneTaq Hot Start 2X Master Mix with Standard Buffer 500 rxns
    Catalog Number:
    m0484l
    Price:
    324
    Size:
    500 rxns
    Category:
    Thermostable DNA Polymerases
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    Structured Review

    New England Biolabs taqman genotyping master mix
    OneTaq Hot Start 2X Master Mix with Standard Buffer
    OneTaq Hot Start 2X Master Mix with Standard Buffer 500 rxns
    https://www.bioz.com/result/taqman genotyping master mix/product/New England Biolabs
    Average 90 stars, based on 18909 article reviews
    Price from $9.99 to $1999.99
    taqman genotyping master mix - by Bioz Stars, 2020-09
    90/100 stars

    Images

    1) Product Images from "Rational “Error Elimination” Approach to Evaluating Molecular Barcoded Next-Generation Sequencing Data Identifies Low-Frequency Mutations in Hematologic Malignancies"

    Article Title: Rational “Error Elimination” Approach to Evaluating Molecular Barcoded Next-Generation Sequencing Data Identifies Low-Frequency Mutations in Hematologic Malignancies

    Journal: The Journal of Molecular Diagnostics : JMD

    doi: 10.1016/j.jmoldx.2019.01.008

    Evaluation of three different polymerase master mixes in the first and second stages of PCR, for sequencing library preparation. Amplification reaction mixes were assembled with TaqMan genotyping master mix, HotStarTaq Plus master mix, or NEBNext Ultra II Q5 mix during first-stage PCR. All of the first-stage PCR products were assembled in NEBNext Ultra II Q5 master mix ( A ), HotStarTaq Plus master mix ( B ), or TaqMan genotyping master mix ( C ) for second-stage PCR. Libraries were purified with solid-phase reversible immobilization beads and analyzed on an Agilent 2100 DNA bioanalyzer. A 300- to 400-bp target-specific library is indicated by a bracket . Note that the fragments of 100 to 200 bp predominantly contained primer dimers. Green and purple bars indicate lower and upper markers, respectively. All samples were evaluated in triplicate.
    Figure Legend Snippet: Evaluation of three different polymerase master mixes in the first and second stages of PCR, for sequencing library preparation. Amplification reaction mixes were assembled with TaqMan genotyping master mix, HotStarTaq Plus master mix, or NEBNext Ultra II Q5 mix during first-stage PCR. All of the first-stage PCR products were assembled in NEBNext Ultra II Q5 master mix ( A ), HotStarTaq Plus master mix ( B ), or TaqMan genotyping master mix ( C ) for second-stage PCR. Libraries were purified with solid-phase reversible immobilization beads and analyzed on an Agilent 2100 DNA bioanalyzer. A 300- to 400-bp target-specific library is indicated by a bracket . Note that the fragments of 100 to 200 bp predominantly contained primer dimers. Green and purple bars indicate lower and upper markers, respectively. All samples were evaluated in triplicate.

    Techniques Used: Polymerase Chain Reaction, Sequencing, Amplification, Purification

    Identification of parameters crucial for improving the quality of molecular barcode–containing next-generation sequencing libraries. A: Exonuclease I treatment reduces the primer dimer concentration and improves the yield of sequencing libraries. First-stage PCR products were incubated with 1 μL of 10 mmol/L Tris-Cl (pH 8.0) or exonuclease I (20 U/μL) at 37°C for 30 minutes. B: Identification of an optimal number of second-stage PCR cycles for library preparation. The first-stage PCR amplification was performed in TaqMan genotyping master mix. The products were then digested with exonuclease I. The second-stage PCR amplification with Ultra II Q5 mix was performed for 17, 20, 23, or 26 cycles. The second-stage PCR products were purified with solid-phase reversible immobilization beads and run on the Agilent 2100 DNA bioanalyzer. C: Size selection efficiently eliminated primer dimers. Genomic DNA mixes A (1% A375, 0.5% Raji, 0.1% NCI-1355, and 98.4% OCI-AML3 DNA; lanes 1, 2, 5, and 6, respectively) and B (1% NCI-1355, 0.5% Raji, 0.1% A375, and 98.4% OCI-AML3 DNA; lanes 3, 4, 7, and 8, respectively) were created and subjected to first-stage PCR amplification, exonuclease I treatment, and second-stage PCR amplification. The purified second-stage PCR products were used for double-size selection with 056×/0.85× volumes of solid-phase reversible immobilization beads, and the size-selected libraries were analyzed on the Agilent 2100 DNA bioanalyzer. Note that a 300- to 400-bp target-specific library is indicated by brackets . Green and purple bars indicate lower and upper markers, respectively. All samples were evaluated in duplicate ( B and C ) or in triplicate ( A ).
    Figure Legend Snippet: Identification of parameters crucial for improving the quality of molecular barcode–containing next-generation sequencing libraries. A: Exonuclease I treatment reduces the primer dimer concentration and improves the yield of sequencing libraries. First-stage PCR products were incubated with 1 μL of 10 mmol/L Tris-Cl (pH 8.0) or exonuclease I (20 U/μL) at 37°C for 30 minutes. B: Identification of an optimal number of second-stage PCR cycles for library preparation. The first-stage PCR amplification was performed in TaqMan genotyping master mix. The products were then digested with exonuclease I. The second-stage PCR amplification with Ultra II Q5 mix was performed for 17, 20, 23, or 26 cycles. The second-stage PCR products were purified with solid-phase reversible immobilization beads and run on the Agilent 2100 DNA bioanalyzer. C: Size selection efficiently eliminated primer dimers. Genomic DNA mixes A (1% A375, 0.5% Raji, 0.1% NCI-1355, and 98.4% OCI-AML3 DNA; lanes 1, 2, 5, and 6, respectively) and B (1% NCI-1355, 0.5% Raji, 0.1% A375, and 98.4% OCI-AML3 DNA; lanes 3, 4, 7, and 8, respectively) were created and subjected to first-stage PCR amplification, exonuclease I treatment, and second-stage PCR amplification. The purified second-stage PCR products were used for double-size selection with 056×/0.85× volumes of solid-phase reversible immobilization beads, and the size-selected libraries were analyzed on the Agilent 2100 DNA bioanalyzer. Note that a 300- to 400-bp target-specific library is indicated by brackets . Green and purple bars indicate lower and upper markers, respectively. All samples were evaluated in duplicate ( B and C ) or in triplicate ( A ).

    Techniques Used: Next-Generation Sequencing, Concentration Assay, Sequencing, Polymerase Chain Reaction, Incubation, Amplification, Purification, Selection

    2) Product Images from "A novel perspective on MOL-PCR optimization and MAGPIX analysis of in-house multiplex foodborne pathogens detection assay"

    Article Title: A novel perspective on MOL-PCR optimization and MAGPIX analysis of in-house multiplex foodborne pathogens detection assay

    Journal: Scientific Reports

    doi: 10.1038/s41598-019-40035-5

    Impact evaluation of Master Mix used in singleplex PCR. In graph: Elizyme = 2X EliZyme HS Robust MIX (Elisabeth Pharmacon, Czech Republic); Accustart II = AccuStart II PCR ToughMix (QuantaBio, Massachusetts, USA); Amplitaq = AmpliTaq Gold 360 Master Mix (Thermo Fisher Scientific, Massachusetts, USA); OneTaq = OneTaq Hot Start 2X Master Mix with GC Buffer (New England BioLabs, Massachusetts, USA); Platinum = Platinum Hot Start PCR 2X Master Mix (Invitrogen, California, USA); HotStarTaq = HotStarTaq DNA Polymerase (Qiagen, Germany). YE = Y . enterocolitica; TG = T . gondii ; plus sign in legend = positive sample; minus sign = NTC.
    Figure Legend Snippet: Impact evaluation of Master Mix used in singleplex PCR. In graph: Elizyme = 2X EliZyme HS Robust MIX (Elisabeth Pharmacon, Czech Republic); Accustart II = AccuStart II PCR ToughMix (QuantaBio, Massachusetts, USA); Amplitaq = AmpliTaq Gold 360 Master Mix (Thermo Fisher Scientific, Massachusetts, USA); OneTaq = OneTaq Hot Start 2X Master Mix with GC Buffer (New England BioLabs, Massachusetts, USA); Platinum = Platinum Hot Start PCR 2X Master Mix (Invitrogen, California, USA); HotStarTaq = HotStarTaq DNA Polymerase (Qiagen, Germany). YE = Y . enterocolitica; TG = T . gondii ; plus sign in legend = positive sample; minus sign = NTC.

    Techniques Used: Polymerase Chain Reaction, Hot Start PCR

    3) Product Images from "Novel Multitarget Real-Time PCR Assay for Rapid Detection of Bordetella Species in Clinical Specimens ▿"

    Article Title: Novel Multitarget Real-Time PCR Assay for Rapid Detection of Bordetella Species in Clinical Specimens ▿

    Journal: Journal of Clinical Microbiology

    doi: 10.1128/JCM.00601-11

    Linear dynamic range of the singleplex and multiplex RT-PCR assays using Bordetella DNAs. (A) IS 481 primers/probe with B. pertussis A639 DNA. •, singleplex assay; the threshold settings for the IS 481 primer/probe set range from 0.05 to 0.15; IS
    Figure Legend Snippet: Linear dynamic range of the singleplex and multiplex RT-PCR assays using Bordetella DNAs. (A) IS 481 primers/probe with B. pertussis A639 DNA. •, singleplex assay; the threshold settings for the IS 481 primer/probe set range from 0.05 to 0.15; IS

    Techniques Used: Multiplex Assay, Reverse Transcription Polymerase Chain Reaction, Singleplex Assay

    Related Articles

    Clone Assay:

    Article Title: The ter Mutation in the Rat Dnd1 Gene Initiates Gonadal Teratomas and Infertility in Both Genders
    Article Snippet: .. Cloning and in vitro protein expression Dnd1ter and Dnd1wt cDNAs were amplified using the OneTaq® Hot Start 2× Master Mix with Standard Buffer (NEB) with primers Dnd1for1 5′-ATGCAGTCCAAACGGGAGTG-′3 and terRev2 5′-TCACTGCTTCACCACAGAAC-3′ at 94°C for 30 sec; 35 cycles: 94°C for 30 sec, 60°C for 20 sec, 68°C for 1 min; 68°C for 5 min. .. The cDNAs were directly cloned into Vivid Colors™ pcDNA™6.2/N-EmGFP-GW/TOPO® Mammalian Expression Vector (Life Technologies, Darmstadt, Germany) for expression of GFP-Dnd1wt or GFP-Dnd1ter fusion proteins, which were verified by sequencing.

    Amplification:

    Article Title: The ter Mutation in the Rat Dnd1 Gene Initiates Gonadal Teratomas and Infertility in Both Genders
    Article Snippet: .. Cloning and in vitro protein expression Dnd1ter and Dnd1wt cDNAs were amplified using the OneTaq® Hot Start 2× Master Mix with Standard Buffer (NEB) with primers Dnd1for1 5′-ATGCAGTCCAAACGGGAGTG-′3 and terRev2 5′-TCACTGCTTCACCACAGAAC-3′ at 94°C for 30 sec; 35 cycles: 94°C for 30 sec, 60°C for 20 sec, 68°C for 1 min; 68°C for 5 min. .. The cDNAs were directly cloned into Vivid Colors™ pcDNA™6.2/N-EmGFP-GW/TOPO® Mammalian Expression Vector (Life Technologies, Darmstadt, Germany) for expression of GFP-Dnd1wt or GFP-Dnd1ter fusion proteins, which were verified by sequencing.

    Article Title: Random-sequence genetic oligomer pools display an innate potential for ligation and recombination
    Article Snippet: .. XNA RT reactions were isolated using streptavidin beads as described previously , prior to amplification using OneTaq hot-start master mix (NEB), or a blend of OneTaq and ThermoScript (Thermo Fisher), with appropriate primers ( ). .. PolyU tailing was performed using 0.2 U/µl polyU polymerase (NEB) in 1X NEB Buffer 2, 0.5 mM UTP for 20 min at 37 °C, followed by heat inactivation for 1 min at 75 °C.

    Article Title: Random-sequence genetic oligomer pools display an innate potential for ligation and recombination
    Article Snippet: .. Next, RT-PCR was performed (with or without prior polyU-tailing) using the SMARTer RT-PCR system (Clontech), according to the manufacturer’s protocol, using the supplied SMARTer oligo as forward primer, and an oligo containing either a polyA- or polyT as reverse primer ( ; ). cDNA was amplified using OneTaq hot-start master mix (NEB) supplemented with appropriate primers ( ). .. DNA synthesis from semi-random XNA reactions.

    In Vitro:

    Article Title: The ter Mutation in the Rat Dnd1 Gene Initiates Gonadal Teratomas and Infertility in Both Genders
    Article Snippet: .. Cloning and in vitro protein expression Dnd1ter and Dnd1wt cDNAs were amplified using the OneTaq® Hot Start 2× Master Mix with Standard Buffer (NEB) with primers Dnd1for1 5′-ATGCAGTCCAAACGGGAGTG-′3 and terRev2 5′-TCACTGCTTCACCACAGAAC-3′ at 94°C for 30 sec; 35 cycles: 94°C for 30 sec, 60°C for 20 sec, 68°C for 1 min; 68°C for 5 min. .. The cDNAs were directly cloned into Vivid Colors™ pcDNA™6.2/N-EmGFP-GW/TOPO® Mammalian Expression Vector (Life Technologies, Darmstadt, Germany) for expression of GFP-Dnd1wt or GFP-Dnd1ter fusion proteins, which were verified by sequencing.

    Isolation:

    Article Title: Random-sequence genetic oligomer pools display an innate potential for ligation and recombination
    Article Snippet: .. XNA RT reactions were isolated using streptavidin beads as described previously , prior to amplification using OneTaq hot-start master mix (NEB), or a blend of OneTaq and ThermoScript (Thermo Fisher), with appropriate primers ( ). .. PolyU tailing was performed using 0.2 U/µl polyU polymerase (NEB) in 1X NEB Buffer 2, 0.5 mM UTP for 20 min at 37 °C, followed by heat inactivation for 1 min at 75 °C.

    Size-exclusion Chromatography:

    Article Title: The ter Mutation in the Rat Dnd1 Gene Initiates Gonadal Teratomas and Infertility in Both Genders
    Article Snippet: .. Cloning and in vitro protein expression Dnd1ter and Dnd1wt cDNAs were amplified using the OneTaq® Hot Start 2× Master Mix with Standard Buffer (NEB) with primers Dnd1for1 5′-ATGCAGTCCAAACGGGAGTG-′3 and terRev2 5′-TCACTGCTTCACCACAGAAC-3′ at 94°C for 30 sec; 35 cycles: 94°C for 30 sec, 60°C for 20 sec, 68°C for 1 min; 68°C for 5 min. .. The cDNAs were directly cloned into Vivid Colors™ pcDNA™6.2/N-EmGFP-GW/TOPO® Mammalian Expression Vector (Life Technologies, Darmstadt, Germany) for expression of GFP-Dnd1wt or GFP-Dnd1ter fusion proteins, which were verified by sequencing.

    Hot Start PCR:

    Article Title: A novel perspective on MOL-PCR optimization and MAGPIX analysis of in-house multiplex foodborne pathogens detection assay
    Article Snippet: .. Several master mixes were tested in the optimization experiments, namely AccuStart II PCR ToughMix (QuantaBio, Massachusetts, USA), AmpliTaq Gold 360 Master Mix (Thermo Fisher Scientific, Massachusetts, USA), OneTaq Hot Start 2X Master Mix with GC Buffer (New England BioLabs, Massachusetts, USA), Platinum Hot Start PCR Master Mix (Invitrogen, California, USA), and HotStarTaq DNA Polymerase (Qiagen, Germany). .. Each singleplex PCR reaction and thermal cycling protocol was run according to the relevant instructions from the manufacturer.

    Polymerase Chain Reaction:

    Article Title: Ericoid mycorrhizal diversity increases with soil age and progressive phosphorus limitation across a 4.1 million-year chronosequence
    Article Snippet: .. PCR was carried out in 25 μl reactions using 1 μl of template DNA (diluted 1:20), 0.5 μl of each 10 μM primer, 12.5 μl OneTaq Hot-Start 2X Master Mix (New England Biolabs) and a cycling program consisting of: initial denaturing at 94 °C (1 min), 30 cycles of 95 °C (30 sec), 52 °C (30 sec) and 68 °C (30 sec) and a final elongation stage of 68 °C (5 min). .. PCR reactions were carried out in triplicate for each sample and were individually checked for successful amplification using gel electrophoresis.

    Article Title: Comparison of T7E1 and Surveyor Mismatch Cleavage Assays to Detect Mutations Triggered by Engineered Nucleases
    Article Snippet: .. Each 20-µL PCR contained 10 µL of OneTaq Hot Start 2X Master Mix (New England Biolabs), 400 nM of each primer and nuclease-free water. .. PCR conditions were 1 cycle of 5 min at 95°, followed by 40 cycles of 30 sec at 95°, 30 sec at 50° or 58° (depending on the primer pairs, see Table S1 ), and 30 sec at 72°, and finishing with 5 min incubation at 72°.

    Article Title: A novel perspective on MOL-PCR optimization and MAGPIX analysis of in-house multiplex foodborne pathogens detection assay
    Article Snippet: .. Several master mixes were tested in the optimization experiments, namely AccuStart II PCR ToughMix (QuantaBio, Massachusetts, USA), AmpliTaq Gold 360 Master Mix (Thermo Fisher Scientific, Massachusetts, USA), OneTaq Hot Start 2X Master Mix with GC Buffer (New England BioLabs, Massachusetts, USA), Platinum Hot Start PCR Master Mix (Invitrogen, California, USA), and HotStarTaq DNA Polymerase (Qiagen, Germany). .. Each singleplex PCR reaction and thermal cycling protocol was run according to the relevant instructions from the manufacturer.

    Expressing:

    Article Title: The ter Mutation in the Rat Dnd1 Gene Initiates Gonadal Teratomas and Infertility in Both Genders
    Article Snippet: .. Cloning and in vitro protein expression Dnd1ter and Dnd1wt cDNAs were amplified using the OneTaq® Hot Start 2× Master Mix with Standard Buffer (NEB) with primers Dnd1for1 5′-ATGCAGTCCAAACGGGAGTG-′3 and terRev2 5′-TCACTGCTTCACCACAGAAC-3′ at 94°C for 30 sec; 35 cycles: 94°C for 30 sec, 60°C for 20 sec, 68°C for 1 min; 68°C for 5 min. .. The cDNAs were directly cloned into Vivid Colors™ pcDNA™6.2/N-EmGFP-GW/TOPO® Mammalian Expression Vector (Life Technologies, Darmstadt, Germany) for expression of GFP-Dnd1wt or GFP-Dnd1ter fusion proteins, which were verified by sequencing.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Random-sequence genetic oligomer pools display an innate potential for ligation and recombination
    Article Snippet: .. Next, RT-PCR was performed (with or without prior polyU-tailing) using the SMARTer RT-PCR system (Clontech), according to the manufacturer’s protocol, using the supplied SMARTer oligo as forward primer, and an oligo containing either a polyA- or polyT as reverse primer ( ; ). cDNA was amplified using OneTaq hot-start master mix (NEB) supplemented with appropriate primers ( ). .. DNA synthesis from semi-random XNA reactions.

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    New England Biolabs nebnext singleplex adaptors
    Nebnext Singleplex Adaptors, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nebnext singleplex adaptors/product/New England Biolabs
    Average 95 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    nebnext singleplex adaptors - by Bioz Stars, 2020-09
    95/100 stars
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