isothermal amplification buffer  (New England Biolabs)


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    Name:
    Isothermal Amplification Buffer
    Description:
    Isothermal Amplification Buffer 6 0 ml
    Catalog Number:
    b0537s
    Price:
    28
    Size:
    6 0 ml
    Category:
    Buffers
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    New England Biolabs isothermal amplification buffer
    Isothermal Amplification Buffer
    Isothermal Amplification Buffer 6 0 ml
    https://www.bioz.com/result/isothermal amplification buffer/product/New England Biolabs
    Average 95 stars, based on 61 article reviews
    Price from $9.99 to $1999.99
    isothermal amplification buffer - by Bioz Stars, 2020-02
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    Amplification:

    Article Title: Development of loop-mediated isothermal amplification (LAMP) for simple detection of Leishmania infection
    Article Snippet: .. Loop-mediated isothermal amplification The LAMP reaction mixtures (25 μl) were based on that described by Tomita et al. [ ], which contained 1× Isothermal Amplification Buffer (New England Biolabs, USA), 8 mM MgSO4 , 0.8 M Betaine (Sigma-Aldrich, USA), 1.4 mM each of dATP, dCTP, dGTP, and dTTP (SibEnzyme, Russia), 40 pmol of FIP primer, 40 pmol of BIP primer, 10 pmol of F3 primer, 10 pmol of B3 primer, and 8 U of Bst 2.0 WarmStart® DNA Polymerase (New England Biolabs, USA). .. In addition, the colorimetric indicator, 0.008 % MG (Sigma-Aldrich, USA) was added.

    Article Title: Development of a loop-mediated isothermal amplification (LAMP) assay for rapid screening of ticks and fleas for spotted fever group rickettsia
    Article Snippet: .. Bst 2.0 LAMP Each LAMP reaction totaled 25 μl consisting of 9 μl nuclease free water, 2.5 μl 10x Isothermal Amplification Buffer Pack (contains 20mM Tris-HCl, 10mM (NH4 )2 SO4 , 50mM KCl, 2mM MgSO4 , 0.1% Tween® 20) (New England Biolabs), 3.5 μl 10 mM each dNTPs (New England Biolabs), 1 μl 100 mM MgSO4 (New England Biolabs), 3.5 μl 5M Betaine (Affymetrix), and 2.5 μl 10x primer mix (2μM F3 primer, 2μM B3 primer, 16μM FIP, 16μM BIP, 8μM LoopF and 8μM LoopB), 1.0 μl Bst 2.0 WarmStart DNA Polymerase (8 units/μl) (New England Biolabs), 1.0 μl 3mM Hydroxynaphthol blue (HNB) (Sigma-Aldrich), and 1 μl of sample DNA. ..

    Article Title: Visual Detection of Bacterial Pathogens via PNA-Based Padlock Probe Assembly and Isothermal Amplification of DNAzymes
    Article Snippet: .. The ELRCA reaction was performed with 100pM of the circular template, 8 units of Bst2.0 DNA Polymerase (obtained from New England Biolabs), 1× Isothermal Amplification Buffer (obtained from New England Biolabs), 1 mM dNTPs, and 1 μM of each primer. .. Limit of Detection Assay A synthetic target template was used to ligate the ODN probe and evaluate the limit of detection.

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: .. Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA. .. Reaction tubes were placed in a heating block at a constant temperature of 63°C for 60 min and then heated at 80°C for 5 min to stop the reaction.

    Article Title: Sensitive Detection of Xanthomonas oryzae Pathovars oryzae and oryzicola by Loop-Mediated Isothermal Amplification
    Article Snippet: .. The 25-μl reaction mix contained 2.5 μl 10× isothermal amplification buffer (New England BioLabs, Ipswich, MA), 1.4 mM deoxynucleoside triphosphates, 6 mM additional MgSO4 for a final MgSO4 concentration of 8 mM (New England BioLabs, Ipswich, MA), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 4 U Bst DNA polymerase large fragment or Bst DNA polymerase 2.0 (New England BioLabs, Ipswich, MA), 0.32 μM primers FIP and BIP, 32 nM primers F3 and B3, and 0.16 μM primers LoopF and LoopB (loop primers in pathovar-specific assays only) with 1 μl of 20 ng μl−1 DNA, heat-killed cells, or plant extract. .. Mineral oil (EMD Millipore, Darmstadt, Germany) was added on top of the reaction mixture (20 μl) to minimize the introduction of aerosolized product into the work spaces.

    Article Title: Integrated LAMP and immunoassay platform for diarrheal disease detection
    Article Snippet: .. Reactions consist of 1 μL of target sample in 9 μL of LAMP reaction mix containing 1× Isothermal Amplification Buffer (20 mM Tris-HCl, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween 20, pH 8.8@25°C) (NEB), 800 mM of Betaine (Sigma), 0.14 mM of each nucleotide, 6 mM MgSO4 , 0.2 μM each F3 and B3, 0.8 μM each LF and LB, 1.6 μM each FIP and BIP, 1.6 μM quencher primers ( , see Supplementary material), and 3.2 units of Bst 2.0 WarmStart DNA Polymerase (NEB). ..

    Article Title: Direct loop mediated isothermal amplification on filters for quantification of Dehalobacter in groundwater
    Article Snippet: .. LAMP reactions consisted of 1× isothermal amplification buffer (New England Biolabs), 1.4 mM each dNTP (Invitrogen), 0.8 M Betaine solution (Sigma Aldrich), 6 mMMgSO4 (New England Biolabs), 8 U Bst Polymerase 2.0 WarmStart (New England Biolabs), 200 μM SYTO82 Orange Fluorescent Nucleic Acid Stain (ThermoFisher Scientific), template that constitutes 10% of the reaction volume (DNA, crudely lysed, or direct cells), and PCR grade water. .. All DNA extractions were performed using the PowerWater DNA Isolation Kit (12888-100, MoBio Laboratories, Inc.).

    Article Title: Massively parallel biophysical analysis of CRISPR-Cas complexes on next generation sequencing chips
    Article Snippet: .. CJ.RP was extended at 60°C for 10 minutes in isothermal amplification buffer (20 mM Tris-HCl, pH 8.8, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween-20) containing 0.08 U/l of Bst 2.0 WarmStart DNA polymerase (New England Biolabs) and 0.8 mM of dNTPs. .. The chip was then washed with 500 l hybridization buffer at 60°C to remove the polymerase (5 minutes).

    Article Title: Electrical Detection of Nucleic Acid Amplification Using an On-Chip Quasi-Reference Electrode and a PVC REFET
    Article Snippet: .. 25 μL of reaction mix consisted of 0.05×–2× Isothermal Amplification Buffer from New England BioLabs, 800 mM Betaine from Sigma-Aldrich, 50 mM KCl, 1.9 μM FIP and BIP primers, 0.24 μM F3 and B3, 0.96 μM Loop-F and Loop-B primers, 1× EvaGreen from Biotium, 6 units of Warmstart Bst 2.0 polymerase from New England BioLabs, 1.3 mM dNTPs from New England BioLabs, 5 mM MgSO4 from Sigma-Aldrich, and template of our targeted E. coli O26 template. .. Optimization of the reaction centered around three major areas: (1) Tris-HCl buffer concentration, (2) starting pH value, and (3) reaction temperature.

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: .. Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA. .. Reaction tubes were placed in a heating block at a constant temperature of 63°C for 60 min and then heated at 80°C for 5 min to stop the reaction.

    Article Title: Ribozyme-catalysed RNA synthesis using triplet building blocks
    Article Snippet: .. Selection library synthesis Round one libraries were synthesised by mutual extension of 4 nmol of oligonucleotides 1baN30 and 1GMPfo or 1GTPfo at 1 μM each in 1 × isothermal amplification buffer (NEB) with 250 μM each dNTP, annealed (80˚C 3 min, 65˚C 5 min) before addition of 0.4 U/μl Bst 2.0 (NEB) and 30 min 65˚C incubation. .. After purification, 375 μg of each DNA (~1.5 × 1015 mole cules) were transcribed in 5 ml transcription reactions (36 mM tris•HCl pH 7.9 (at 25˚C), 1.8 mM spermidine, 9 mM DTT, 10.8 mM MgCl2 , 2 mM each NTP, 1% 10 × MegaShortScript buffer, 2% 1:9 MegaShortScript:NEB T7 RNA polymerase, 37˚C overnight).

    Article Title: A Loop-Mediated Isothermal Amplification (LAMP) Assay for Early Detection of Schistosoma mansoni in Stool Samples: A Diagnostic Approach in a Murine Model
    Article Snippet: .. Thus, LAMP reactions mixtures (25 µL) contained 40 pmol of each FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Bioron), 1× Isothermal Amplification Buffer −20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), betaine (ranging 0.8, 1, 1.2, 1.4 or 1.6 M) (Sigma, USA), supplementary MgSO4 (ranging 2, 4, 6 or 8 mM) (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 µL of template DNA. .. To establish the standard protocol for the two LAMP reaction mixtures assayed, different temperatures were tested using a heating block (K Dry-Bath) set at 61, 63 and 65°C for 60 min and then heated at 80°C for 5 min to terminate the reaction.

    Article Title: A smartphone-based diagnostic platform for rapid detection of Zika, chikungunya, and dengue viruses
    Article Snippet: .. The fresh reaction mixture contained 1x Isothermal Amplification Buffer (20 mM Tris-HCl, 10 mM (NH4 )2SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB), 700 mM of Betaine (Sigma), 0.14 mM of each nucleotide, 8 mM MgSO4 , 2 μM SYTO® 9 Stain (Thermo Fisher Scientific), 2 units of Porcine RNase Inhibitor , 0.2 μM each F3 and B3, 0.8 μM each LF and LB, 1.6 μM each FIP and BIP primers , 3.2 units of avian myeloblastosis virus (AMV) reverse transcriptase (Life Sciences Advanced Technologies Inc., St Petersburg, FL), and 3.2 units of Bst 2.0 WarmStart® DNA Polymerase (NEB). .. To make the dried reagent formulation, isothermal amplification buffer, betaine and magnesium sulfate were removed.

    Article Title: A Fully Integrated In Vitro Diagnostic Microsystem for Pathogen Detection Developed Using a “3D Extensible” Microfluidic Design Paradigm
    Article Snippet: .. A 25 μL LAMP mixture contained 1.6 μM each of the inner primer (FIP and BIP), 0.2 μM each of the outer primer (F3 and B3), 0.4 μM of the loop primer (LF), 1× Isothermal Amplification Buffer (20 mM tris-HCl, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB, Ipswich, MA, USA), 6.0 mM of MgSO4 , 1.4 mM of dNTPs (Sangon Biotech, Shanghai, China), 0.5 M of betaine (Sigma-Aldrich, St. Louis, MO, USA), 0.15 mM of calcein (Coolaber, Beijing, China), 0.5 mM of MnCl2 (Tiandz, Beijing, China), 8 units of Bst 2.0 WarmStart® DNA Polymerase (NEB, Ipswich, MA, USA), 6 μM Bovine serum albumin (BSA) (Sigma-Aldrich, St. Louis, MO, USA), and the template. ..

    Construct:

    Article Title: Ribozyme-catalysed RNA synthesis using triplet building blocks
    Article Snippet: Selection library synthesis Round one libraries were synthesised by mutual extension of 4 nmol of oligonucleotides 1baN30 and 1GMPfo or 1GTPfo at 1 μM each in 1 × isothermal amplification buffer (NEB) with 250 μM each dNTP, annealed (80˚C 3 min, 65˚C 5 min) before addition of 0.4 U/μl Bst 2.0 (NEB) and 30 min 65˚C incubation. .. These were treated with DNase, acid phenol/chloroform extracted and 73% ethanol precipitated prior to urea-PAGE purification, elution, filtering (Spin-X) and re-precipitation, yielding the 1GTP Zcore selection construct ( ).

    SYBR Green Assay:

    Article Title: Direct and Rapid Detection and Quantification of Oenococcus oeni Cells in Wine by Cells-LAMP and Cells-qLAMP
    Article Snippet: .. Each reaction contained 1.4 mM of dNTPs, 0.2 μM of each outer primer, 1.6 μM of each inner primer, 0.8 μM of each loop primer, 8 mM of MgCl2 (Invitrogen), 1× Isothermal Amplification Buffer, 0.4 U/μL of Bst polymerase 2.0 (New England BioLabs), 11.5 μL of the mechanically lysed cells, and 0.4 μM of SYTO-9 (Invitrogen) or 1 μL of 10× SYBR Green I (Invitrogen). .. The qLAMP amplifications were performed in triplicate in a C100TM Thermal Cycler, CFX96TM Real-Time System (BioRad), which operated at a constant temperature of 62°C for 1 h. Fluorescence signals were collected every minute, followed by a melting curve analysis obtained by slow heating from 60°C to 95°C at 0.5°C every 5 s, with continuous fluorescence collection.

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA. .. The LAMP-positive results could be visually inspected by naked eye by color change after adding 2 μL of 1:10 diluted 10,000x concentration fluorescent dye SYBR Green I to the reactions tubes.

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA. .. The LAMP-positive results could be visually inspected by naked eye by color change after adding 2 μL of 1:10 diluted 10,000x concentration fluorescent dye SYBR Green I to the reactions tubes.

    Incubation:

    Article Title: Sensitive Detection of Xanthomonas oryzae Pathovars oryzae and oryzicola by Loop-Mediated Isothermal Amplification
    Article Snippet: The 25-μl reaction mix contained 2.5 μl 10× isothermal amplification buffer (New England BioLabs, Ipswich, MA), 1.4 mM deoxynucleoside triphosphates, 6 mM additional MgSO4 for a final MgSO4 concentration of 8 mM (New England BioLabs, Ipswich, MA), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 4 U Bst DNA polymerase large fragment or Bst DNA polymerase 2.0 (New England BioLabs, Ipswich, MA), 0.32 μM primers FIP and BIP, 32 nM primers F3 and B3, and 0.16 μM primers LoopF and LoopB (loop primers in pathovar-specific assays only) with 1 μl of 20 ng μl−1 DNA, heat-killed cells, or plant extract. .. Amplification was terminated by heat inactivation at 80°C for 3 min. After incubation, the tubes were individually opened in a separate lab and 0.5 to 1 μl of Quant-IT Pico green reagent (Invitrogen, Carlsbad, CA) was added.

    Article Title: Direct loop mediated isothermal amplification on filters for quantification of Dehalobacter in groundwater
    Article Snippet: LAMP reactions consisted of 1× isothermal amplification buffer (New England Biolabs), 1.4 mM each dNTP (Invitrogen), 0.8 M Betaine solution (Sigma Aldrich), 6 mMMgSO4 (New England Biolabs), 8 U Bst Polymerase 2.0 WarmStart (New England Biolabs), 200 μM SYTO82 Orange Fluorescent Nucleic Acid Stain (ThermoFisher Scientific), template that constitutes 10% of the reaction volume (DNA, crudely lysed, or direct cells), and PCR grade water. .. All experiments were performed with an isothermal incubation at 63 °C for 60 min with plate reads at one minute intervals in the real-time thermal cycler, and every 16 s in the Gene-Z device.

    Article Title: Massively parallel biophysical analysis of CRISPR-Cas complexes on next generation sequencing chips
    Article Snippet: After denaturation, the chip was heated to 85°C and incubated with 500 nM of the regeneration primer (CJ.RP) in hybridization buffer (75 mM Trisodium Citrate, pH 7.0, 750 mM NaCl, 0.1% Tween-20). .. CJ.RP was extended at 60°C for 10 minutes in isothermal amplification buffer (20 mM Tris-HCl, pH 8.8, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween-20) containing 0.08 U/l of Bst 2.0 WarmStart DNA polymerase (New England Biolabs) and 0.8 mM of dNTPs.

    Article Title: Ribozyme-catalysed RNA synthesis using triplet building blocks
    Article Snippet: .. Selection library synthesis Round one libraries were synthesised by mutual extension of 4 nmol of oligonucleotides 1baN30 and 1GMPfo or 1GTPfo at 1 μM each in 1 × isothermal amplification buffer (NEB) with 250 μM each dNTP, annealed (80˚C 3 min, 65˚C 5 min) before addition of 0.4 U/μl Bst 2.0 (NEB) and 30 min 65˚C incubation. .. After purification, 375 μg of each DNA (~1.5 × 1015 mole cules) were transcribed in 5 ml transcription reactions (36 mM tris•HCl pH 7.9 (at 25˚C), 1.8 mM spermidine, 9 mM DTT, 10.8 mM MgCl2 , 2 mM each NTP, 1% 10 × MegaShortScript buffer, 2% 1:9 MegaShortScript:NEB T7 RNA polymerase, 37˚C overnight).

    Article Title: A smartphone-based diagnostic platform for rapid detection of Zika, chikungunya, and dengue viruses
    Article Snippet: The fresh reaction mixture contained 1x Isothermal Amplification Buffer (20 mM Tris-HCl, 10 mM (NH4 )2SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB), 700 mM of Betaine (Sigma), 0.14 mM of each nucleotide, 8 mM MgSO4 , 2 μM SYTO® 9 Stain (Thermo Fisher Scientific), 2 units of Porcine RNase Inhibitor , 0.2 μM each F3 and B3, 0.8 μM each LF and LB, 1.6 μM each FIP and BIP primers , 3.2 units of avian myeloblastosis virus (AMV) reverse transcriptase (Life Sciences Advanced Technologies Inc., St Petersburg, FL), and 3.2 units of Bst 2.0 WarmStart® DNA Polymerase (NEB). .. For real-time monitoring of the reaction, fluorescent intensity was collected every 1 minute through FAM channel during incubation at 67 °C for 40 minutes and for the end-point, fluorescent intensity was measured at room temperature though Cy5 channel after the incubation using CFX-96 real-time thermocycler (Bio-Rad Laboratories, Hercules, CA).

    Hybridization:

    Article Title: Massively parallel biophysical analysis of CRISPR-Cas complexes on next generation sequencing chips
    Article Snippet: After denaturation, the chip was heated to 85°C and incubated with 500 nM of the regeneration primer (CJ.RP) in hybridization buffer (75 mM Trisodium Citrate, pH 7.0, 750 mM NaCl, 0.1% Tween-20). .. CJ.RP was extended at 60°C for 10 minutes in isothermal amplification buffer (20 mM Tris-HCl, pH 8.8, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween-20) containing 0.08 U/l of Bst 2.0 WarmStart DNA polymerase (New England Biolabs) and 0.8 mM of dNTPs.

    Sequencing:

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: The SmMIT-LAMP method amplifies a specific sequence corresponding to a mitochondrial S . mansoni minisatellite DNA region (GenBank Acc. .. Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA.

    Article Title: Massively parallel biophysical analysis of CRISPR-Cas complexes on next generation sequencing chips
    Article Snippet: This removed the untethered DNAs strands containing residual fluorescent dyes from sequencing (see ). .. CJ.RP was extended at 60°C for 10 minutes in isothermal amplification buffer (20 mM Tris-HCl, pH 8.8, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween-20) containing 0.08 U/l of Bst 2.0 WarmStart DNA polymerase (New England Biolabs) and 0.8 mM of dNTPs.

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: The SmMIT-LAMP method amplifies a specific sequence corresponding to a mitochondrial S . mansoni minisatellite DNA region (GenBank Acc. .. Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA.

    Article Title: A Fully Integrated In Vitro Diagnostic Microsystem for Pathogen Detection Developed Using a “3D Extensible” Microfluidic Design Paradigm
    Article Snippet: The detection of Chlamydia trachomatis (CT) was realized by amplifying a specific sequence in its 7.5 kb cryptic plasmid. .. A 25 μL LAMP mixture contained 1.6 μM each of the inner primer (FIP and BIP), 0.2 μM each of the outer primer (F3 and B3), 0.4 μM of the loop primer (LF), 1× Isothermal Amplification Buffer (20 mM tris-HCl, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB, Ipswich, MA, USA), 6.0 mM of MgSO4 , 1.4 mM of dNTPs (Sangon Biotech, Shanghai, China), 0.5 M of betaine (Sigma-Aldrich, St. Louis, MO, USA), 0.15 mM of calcein (Coolaber, Beijing, China), 0.5 mM of MnCl2 (Tiandz, Beijing, China), 8 units of Bst 2.0 WarmStart® DNA Polymerase (NEB, Ipswich, MA, USA), 6 μM Bovine serum albumin (BSA) (Sigma-Aldrich, St. Louis, MO, USA), and the template.

    DNA Extraction:

    Article Title: Direct loop mediated isothermal amplification on filters for quantification of Dehalobacter in groundwater
    Article Snippet: LAMP reactions consisted of 1× isothermal amplification buffer (New England Biolabs), 1.4 mM each dNTP (Invitrogen), 0.8 M Betaine solution (Sigma Aldrich), 6 mMMgSO4 (New England Biolabs), 8 U Bst Polymerase 2.0 WarmStart (New England Biolabs), 200 μM SYTO82 Orange Fluorescent Nucleic Acid Stain (ThermoFisher Scientific), template that constitutes 10% of the reaction volume (DNA, crudely lysed, or direct cells), and PCR grade water. .. All DNA extractions were performed using the PowerWater DNA Isolation Kit (12888-100, MoBio Laboratories, Inc.).

    Article Title: A Fully Integrated In Vitro Diagnostic Microsystem for Pathogen Detection Developed Using a “3D Extensible” Microfluidic Design Paradigm
    Article Snippet: Paragraph title: 2.4. DNA Extraction and Loop-Mediated Isothermal Amplification (LAMP) Reaction ... A 25 μL LAMP mixture contained 1.6 μM each of the inner primer (FIP and BIP), 0.2 μM each of the outer primer (F3 and B3), 0.4 μM of the loop primer (LF), 1× Isothermal Amplification Buffer (20 mM tris-HCl, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB, Ipswich, MA, USA), 6.0 mM of MgSO4 , 1.4 mM of dNTPs (Sangon Biotech, Shanghai, China), 0.5 M of betaine (Sigma-Aldrich, St. Louis, MO, USA), 0.15 mM of calcein (Coolaber, Beijing, China), 0.5 mM of MnCl2 (Tiandz, Beijing, China), 8 units of Bst 2.0 WarmStart® DNA Polymerase (NEB, Ipswich, MA, USA), 6 μM Bovine serum albumin (BSA) (Sigma-Aldrich, St. Louis, MO, USA), and the template.

    Fluorescence:

    Article Title: Direct and Rapid Detection and Quantification of Oenococcus oeni Cells in Wine by Cells-LAMP and Cells-qLAMP
    Article Snippet: Each reaction contained 1.4 mM of dNTPs, 0.2 μM of each outer primer, 1.6 μM of each inner primer, 0.8 μM of each loop primer, 8 mM of MgCl2 (Invitrogen), 1× Isothermal Amplification Buffer, 0.4 U/μL of Bst polymerase 2.0 (New England BioLabs), 11.5 μL of the mechanically lysed cells, and 0.4 μM of SYTO-9 (Invitrogen) or 1 μL of 10× SYBR Green I (Invitrogen). .. The qLAMP amplifications were performed in triplicate in a C100TM Thermal Cycler, CFX96TM Real-Time System (BioRad), which operated at a constant temperature of 62°C for 1 h. Fluorescence signals were collected every minute, followed by a melting curve analysis obtained by slow heating from 60°C to 95°C at 0.5°C every 5 s, with continuous fluorescence collection.

    Article Title: Sensitive Detection of Xanthomonas oryzae Pathovars oryzae and oryzicola by Loop-Mediated Isothermal Amplification
    Article Snippet: The 25-μl reaction mix contained 2.5 μl 10× isothermal amplification buffer (New England BioLabs, Ipswich, MA), 1.4 mM deoxynucleoside triphosphates, 6 mM additional MgSO4 for a final MgSO4 concentration of 8 mM (New England BioLabs, Ipswich, MA), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 4 U Bst DNA polymerase large fragment or Bst DNA polymerase 2.0 (New England BioLabs, Ipswich, MA), 0.32 μM primers FIP and BIP, 32 nM primers F3 and B3, and 0.16 μM primers LoopF and LoopB (loop primers in pathovar-specific assays only) with 1 μl of 20 ng μl−1 DNA, heat-killed cells, or plant extract. .. The reaction mixtures were incubated at room temperature for 5 min and then observed under normal and UV light either for a change of color from orange to green or for fluorescence.

    RT Lamp Assay:

    Article Title: A smartphone-based diagnostic platform for rapid detection of Zika, chikungunya, and dengue viruses
    Article Snippet: Paragraph title: RT-LAMP assay ... The fresh reaction mixture contained 1x Isothermal Amplification Buffer (20 mM Tris-HCl, 10 mM (NH4 )2SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB), 700 mM of Betaine (Sigma), 0.14 mM of each nucleotide, 8 mM MgSO4 , 2 μM SYTO® 9 Stain (Thermo Fisher Scientific), 2 units of Porcine RNase Inhibitor , 0.2 μM each F3 and B3, 0.8 μM each LF and LB, 1.6 μM each FIP and BIP primers , 3.2 units of avian myeloblastosis virus (AMV) reverse transcriptase (Life Sciences Advanced Technologies Inc., St Petersburg, FL), and 3.2 units of Bst 2.0 WarmStart® DNA Polymerase (NEB).

    Labeling:

    Article Title: Massively parallel biophysical analysis of CRISPR-Cas complexes on next generation sequencing chips
    Article Snippet: CJ.RP was extended at 60°C for 10 minutes in isothermal amplification buffer (20 mM Tris-HCl, pH 8.8, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween-20) containing 0.08 U/l of Bst 2.0 WarmStart DNA polymerase (New England Biolabs) and 0.8 mM of dNTPs. .. Finally, a phiX primer labeled with Atto647 or Cy3 (atto647-PCP/Cy3-PCP) was annealed under the same conditions as CJ.RP.

    Purification:

    Article Title: Ribozyme-catalysed RNA synthesis using triplet building blocks
    Article Snippet: Selection library synthesis Round one libraries were synthesised by mutual extension of 4 nmol of oligonucleotides 1baN30 and 1GMPfo or 1GTPfo at 1 μM each in 1 × isothermal amplification buffer (NEB) with 250 μM each dNTP, annealed (80˚C 3 min, 65˚C 5 min) before addition of 0.4 U/μl Bst 2.0 (NEB) and 30 min 65˚C incubation. .. After purification, 375 μg of each DNA (~1.5 × 1015 mole cules) were transcribed in 5 ml transcription reactions (36 mM tris•HCl pH 7.9 (at 25˚C), 1.8 mM spermidine, 9 mM DTT, 10.8 mM MgCl2 , 2 mM each NTP, 1% 10 × MegaShortScript buffer, 2% 1:9 MegaShortScript:NEB T7 RNA polymerase, 37˚C overnight).

    Polymerase Chain Reaction:

    Article Title: Direct loop mediated isothermal amplification on filters for quantification of Dehalobacter in groundwater
    Article Snippet: .. LAMP reactions consisted of 1× isothermal amplification buffer (New England Biolabs), 1.4 mM each dNTP (Invitrogen), 0.8 M Betaine solution (Sigma Aldrich), 6 mMMgSO4 (New England Biolabs), 8 U Bst Polymerase 2.0 WarmStart (New England Biolabs), 200 μM SYTO82 Orange Fluorescent Nucleic Acid Stain (ThermoFisher Scientific), template that constitutes 10% of the reaction volume (DNA, crudely lysed, or direct cells), and PCR grade water. .. All DNA extractions were performed using the PowerWater DNA Isolation Kit (12888-100, MoBio Laboratories, Inc.).

    Article Title: A smartphone-based diagnostic platform for rapid detection of Zika, chikungunya, and dengue viruses
    Article Snippet: RT-LAMP assay RT-LAMP reactions were carried out in PCR tubes or 96-well plates with 1 μL of the sample in 9 μL of RT-LAMP reaction solution. .. The fresh reaction mixture contained 1x Isothermal Amplification Buffer (20 mM Tris-HCl, 10 mM (NH4 )2SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB), 700 mM of Betaine (Sigma), 0.14 mM of each nucleotide, 8 mM MgSO4 , 2 μM SYTO® 9 Stain (Thermo Fisher Scientific), 2 units of Porcine RNase Inhibitor , 0.2 μM each F3 and B3, 0.8 μM each LF and LB, 1.6 μM each FIP and BIP primers , 3.2 units of avian myeloblastosis virus (AMV) reverse transcriptase (Life Sciences Advanced Technologies Inc., St Petersburg, FL), and 3.2 units of Bst 2.0 WarmStart® DNA Polymerase (NEB).

    Blocking Assay:

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA. .. Reaction tubes were placed in a heating block at a constant temperature of 63°C for 60 min and then heated at 80°C for 5 min to stop the reaction.

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA. .. Reaction tubes were placed in a heating block at a constant temperature of 63°C for 60 min and then heated at 80°C for 5 min to stop the reaction.

    Article Title: A Loop-Mediated Isothermal Amplification (LAMP) Assay for Early Detection of Schistosoma mansoni in Stool Samples: A Diagnostic Approach in a Murine Model
    Article Snippet: Thus, LAMP reactions mixtures (25 µL) contained 40 pmol of each FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Bioron), 1× Isothermal Amplification Buffer −20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), betaine (ranging 0.8, 1, 1.2, 1.4 or 1.6 M) (Sigma, USA), supplementary MgSO4 (ranging 2, 4, 6 or 8 mM) (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 µL of template DNA. .. To establish the standard protocol for the two LAMP reaction mixtures assayed, different temperatures were tested using a heating block (K Dry-Bath) set at 61, 63 and 65°C for 60 min and then heated at 80°C for 5 min to terminate the reaction.

    Staining:

    Article Title: Direct loop mediated isothermal amplification on filters for quantification of Dehalobacter in groundwater
    Article Snippet: .. LAMP reactions consisted of 1× isothermal amplification buffer (New England Biolabs), 1.4 mM each dNTP (Invitrogen), 0.8 M Betaine solution (Sigma Aldrich), 6 mMMgSO4 (New England Biolabs), 8 U Bst Polymerase 2.0 WarmStart (New England Biolabs), 200 μM SYTO82 Orange Fluorescent Nucleic Acid Stain (ThermoFisher Scientific), template that constitutes 10% of the reaction volume (DNA, crudely lysed, or direct cells), and PCR grade water. .. All DNA extractions were performed using the PowerWater DNA Isolation Kit (12888-100, MoBio Laboratories, Inc.).

    Article Title: A smartphone-based diagnostic platform for rapid detection of Zika, chikungunya, and dengue viruses
    Article Snippet: .. The fresh reaction mixture contained 1x Isothermal Amplification Buffer (20 mM Tris-HCl, 10 mM (NH4 )2SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB), 700 mM of Betaine (Sigma), 0.14 mM of each nucleotide, 8 mM MgSO4 , 2 μM SYTO® 9 Stain (Thermo Fisher Scientific), 2 units of Porcine RNase Inhibitor , 0.2 μM each F3 and B3, 0.8 μM each LF and LB, 1.6 μM each FIP and BIP primers , 3.2 units of avian myeloblastosis virus (AMV) reverse transcriptase (Life Sciences Advanced Technologies Inc., St Petersburg, FL), and 3.2 units of Bst 2.0 WarmStart® DNA Polymerase (NEB). .. To make the dried reagent formulation, isothermal amplification buffer, betaine and magnesium sulfate were removed.

    Chromatin Immunoprecipitation:

    Article Title: Massively parallel biophysical analysis of CRISPR-Cas complexes on next generation sequencing chips
    Article Snippet: Paragraph title: Chip regeneration and addition of alignment markers ... CJ.RP was extended at 60°C for 10 minutes in isothermal amplification buffer (20 mM Tris-HCl, pH 8.8, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween-20) containing 0.08 U/l of Bst 2.0 WarmStart DNA polymerase (New England Biolabs) and 0.8 mM of dNTPs.

    Plasmid Preparation:

    Article Title: A Fully Integrated In Vitro Diagnostic Microsystem for Pathogen Detection Developed Using a “3D Extensible” Microfluidic Design Paradigm
    Article Snippet: Each CT has 7~10 copies of this plasmid and the sequences of the template and LAMP primers are listed in . .. A 25 μL LAMP mixture contained 1.6 μM each of the inner primer (FIP and BIP), 0.2 μM each of the outer primer (F3 and B3), 0.4 μM of the loop primer (LF), 1× Isothermal Amplification Buffer (20 mM tris-HCl, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB, Ipswich, MA, USA), 6.0 mM of MgSO4 , 1.4 mM of dNTPs (Sangon Biotech, Shanghai, China), 0.5 M of betaine (Sigma-Aldrich, St. Louis, MO, USA), 0.15 mM of calcein (Coolaber, Beijing, China), 0.5 mM of MnCl2 (Tiandz, Beijing, China), 8 units of Bst 2.0 WarmStart® DNA Polymerase (NEB, Ipswich, MA, USA), 6 μM Bovine serum albumin (BSA) (Sigma-Aldrich, St. Louis, MO, USA), and the template.

    Real-time Polymerase Chain Reaction:

    Article Title: Direct loop mediated isothermal amplification on filters for quantification of Dehalobacter in groundwater
    Article Snippet: Paragraph title: 2.2. qPCR and LAMP experiments ... LAMP reactions consisted of 1× isothermal amplification buffer (New England Biolabs), 1.4 mM each dNTP (Invitrogen), 0.8 M Betaine solution (Sigma Aldrich), 6 mMMgSO4 (New England Biolabs), 8 U Bst Polymerase 2.0 WarmStart (New England Biolabs), 200 μM SYTO82 Orange Fluorescent Nucleic Acid Stain (ThermoFisher Scientific), template that constitutes 10% of the reaction volume (DNA, crudely lysed, or direct cells), and PCR grade water.

    Article Title: Electrical Detection of Nucleic Acid Amplification Using an On-Chip Quasi-Reference Electrode and a PVC REFET
    Article Snippet: 25 μL of reaction mix consisted of 0.05×–2× Isothermal Amplification Buffer from New England BioLabs, 800 mM Betaine from Sigma-Aldrich, 50 mM KCl, 1.9 μM FIP and BIP primers, 0.24 μM F3 and B3, 0.96 μM Loop-F and Loop-B primers, 1× EvaGreen from Biotium, 6 units of Warmstart Bst 2.0 polymerase from New England BioLabs, 1.3 mM dNTPs from New England BioLabs, 5 mM MgSO4 from Sigma-Aldrich, and template of our targeted E. coli O26 template. .. Electrical detection of LAMP was carried out by first running part of the solution in an Eppendorf RealPlex qPCR system.

    Selection:

    Article Title: Ribozyme-catalysed RNA synthesis using triplet building blocks
    Article Snippet: .. Selection library synthesis Round one libraries were synthesised by mutual extension of 4 nmol of oligonucleotides 1baN30 and 1GMPfo or 1GTPfo at 1 μM each in 1 × isothermal amplification buffer (NEB) with 250 μM each dNTP, annealed (80˚C 3 min, 65˚C 5 min) before addition of 0.4 U/μl Bst 2.0 (NEB) and 30 min 65˚C incubation. .. After purification, 375 μg of each DNA (~1.5 × 1015 mole cules) were transcribed in 5 ml transcription reactions (36 mM tris•HCl pH 7.9 (at 25˚C), 1.8 mM spermidine, 9 mM DTT, 10.8 mM MgCl2 , 2 mM each NTP, 1% 10 × MegaShortScript buffer, 2% 1:9 MegaShortScript:NEB T7 RNA polymerase, 37˚C overnight).

    Agarose Gel Electrophoresis:

    Article Title: Development of loop-mediated isothermal amplification (LAMP) for simple detection of Leishmania infection
    Article Snippet: Loop-mediated isothermal amplification The LAMP reaction mixtures (25 μl) were based on that described by Tomita et al. [ ], which contained 1× Isothermal Amplification Buffer (New England Biolabs, USA), 8 mM MgSO4 , 0.8 M Betaine (Sigma-Aldrich, USA), 1.4 mM each of dATP, dCTP, dGTP, and dTTP (SibEnzyme, Russia), 40 pmol of FIP primer, 40 pmol of BIP primer, 10 pmol of F3 primer, 10 pmol of B3 primer, and 8 U of Bst 2.0 WarmStart® DNA Polymerase (New England Biolabs, USA). .. Visualization of the LAMP products was performed using 2.5 % agarose gel electrophoresis at 10 V/cm in 1× TAE buffer.

    Concentration Assay:

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA. .. The LAMP-positive results could be visually inspected by naked eye by color change after adding 2 μL of 1:10 diluted 10,000x concentration fluorescent dye SYBR Green I to the reactions tubes.

    Article Title: Sensitive Detection of Xanthomonas oryzae Pathovars oryzae and oryzicola by Loop-Mediated Isothermal Amplification
    Article Snippet: .. The 25-μl reaction mix contained 2.5 μl 10× isothermal amplification buffer (New England BioLabs, Ipswich, MA), 1.4 mM deoxynucleoside triphosphates, 6 mM additional MgSO4 for a final MgSO4 concentration of 8 mM (New England BioLabs, Ipswich, MA), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 4 U Bst DNA polymerase large fragment or Bst DNA polymerase 2.0 (New England BioLabs, Ipswich, MA), 0.32 μM primers FIP and BIP, 32 nM primers F3 and B3, and 0.16 μM primers LoopF and LoopB (loop primers in pathovar-specific assays only) with 1 μl of 20 ng μl−1 DNA, heat-killed cells, or plant extract. .. Mineral oil (EMD Millipore, Darmstadt, Germany) was added on top of the reaction mixture (20 μl) to minimize the introduction of aerosolized product into the work spaces.

    Article Title: Electrical Detection of Nucleic Acid Amplification Using an On-Chip Quasi-Reference Electrode and a PVC REFET
    Article Snippet: 25 μL of reaction mix consisted of 0.05×–2× Isothermal Amplification Buffer from New England BioLabs, 800 mM Betaine from Sigma-Aldrich, 50 mM KCl, 1.9 μM FIP and BIP primers, 0.24 μM F3 and B3, 0.96 μM Loop-F and Loop-B primers, 1× EvaGreen from Biotium, 6 units of Warmstart Bst 2.0 polymerase from New England BioLabs, 1.3 mM dNTPs from New England BioLabs, 5 mM MgSO4 from Sigma-Aldrich, and template of our targeted E. coli O26 template. .. Optimization of the reaction centered around three major areas: (1) Tris-HCl buffer concentration, (2) starting pH value, and (3) reaction temperature.

    Article Title: A field survey using LAMP assay for detection of Schistosoma mansoni in a low-transmission area of schistosomiasis in Umbuzeiro, Brazil: Assessment in human and snail samples
    Article Snippet: Briefly, the reaction was carried out in 25 μL reaction mixture containing 40 pmol each of FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Intron), 1x Isothermal Amplification Buffer -20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), 1 M betaine (Sigma, USA), supplementary 6 mM of MgSO4 (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 μL of template DNA. .. The LAMP-positive results could be visually inspected by naked eye by color change after adding 2 μL of 1:10 diluted 10,000x concentration fluorescent dye SYBR Green I to the reactions tubes.

    Lamp Assay:

    Article Title: A Loop-Mediated Isothermal Amplification (LAMP) Assay for Early Detection of Schistosoma mansoni in Stool Samples: A Diagnostic Approach in a Murine Model
    Article Snippet: Paragraph title: LAMP assay ... Thus, LAMP reactions mixtures (25 µL) contained 40 pmol of each FIP and BIP primers, 5 pmol of each F3 and B3 primers, 1.4 mM of each dNTP (Bioron), 1× Isothermal Amplification Buffer −20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 , 0.1% Tween20- (New England Biolabs, UK), betaine (ranging 0.8, 1, 1.2, 1.4 or 1.6 M) (Sigma, USA), supplementary MgSO4 (ranging 2, 4, 6 or 8 mM) (New England Biolabs, UK) and 8 U of Bst 2.0 WarmStart DNA polymerase (New England Biolabs, UK) with 2 µL of template DNA.

    Lysis:

    Article Title: A Fully Integrated In Vitro Diagnostic Microsystem for Pathogen Detection Developed Using a “3D Extensible” Microfluidic Design Paradigm
    Article Snippet: Since 250 μL of lysis buffer was employed to flush the swab, a total of 2500, 250, and 25 CT particles on the swabs can theoretically generate the lysates with concentrations of 10, 1, and 0.1 CT particles/μL, respectively. .. A 25 μL LAMP mixture contained 1.6 μM each of the inner primer (FIP and BIP), 0.2 μM each of the outer primer (F3 and B3), 0.4 μM of the loop primer (LF), 1× Isothermal Amplification Buffer (20 mM tris-HCl, 10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, pH 8.8@25 °C, NEB, Ipswich, MA, USA), 6.0 mM of MgSO4 , 1.4 mM of dNTPs (Sangon Biotech, Shanghai, China), 0.5 M of betaine (Sigma-Aldrich, St. Louis, MO, USA), 0.15 mM of calcein (Coolaber, Beijing, China), 0.5 mM of MnCl2 (Tiandz, Beijing, China), 8 units of Bst 2.0 WarmStart® DNA Polymerase (NEB, Ipswich, MA, USA), 6 μM Bovine serum albumin (BSA) (Sigma-Aldrich, St. Louis, MO, USA), and the template.

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    New England Biolabs isothermal buffer
    Isothermal Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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