warmstart reverse transcriptase  (New England Biolabs)


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    Name:
    WarmStart RTx Reverse Transcriptase
    Description:
    WarmStart RTx Reverse Transcriptase 250 rxns
    Catalog Number:
    m0380l
    Price:
    266
    Size:
    250 rxns
    Category:
    Reverse Transcriptases
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    New England Biolabs warmstart reverse transcriptase
    WarmStart RTx Reverse Transcriptase
    WarmStart RTx Reverse Transcriptase 250 rxns
    https://www.bioz.com/result/warmstart reverse transcriptase/product/New England Biolabs
    Average 95 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    warmstart reverse transcriptase - by Bioz Stars, 2020-02
    95/100 stars

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    Produced:

    Article Title: Field diagnosis and genotyping of chikungunya virus using a dried reverse transcription loop-mediated isothermal amplification (LAMP) assay and MinION sequencing
    Article Snippet: Preparation of in-house dried CHIKV RT-LAMP (CHIKV-CZC-LAMP) system The in-house dried CHIKV RT-LAMP system was produced by using a trehalose vitrification technique based on a previous report [ ] with several modifications. .. The Trehalose solution (1.6 μl, 2M), deoxyribonucleotide triphosphates (dNTPs) (1.4 μl, 25mM each) (Nippon Gene, Tokyo, Japan), WarmStart RTx reverse transcriptase (0.25 μl, 15 U/μl) (New England Biolabs Inc., Ipswich, MA), RNase inhibitor (0.1 μl, 40 U/μl) (Takara Bio Inc., Shiga, Japan), and Bst 2.0 WarmStart DNA polymerase (0.05 μl, 120 U/μl and 0.25 μl, 8 U/μl) (New England Biolabs Inc.) were then mixed.

    Multiplex Assay:

    Article Title: Simultaneous Detection of Different Zika Virus Lineages via Molecular Computation in a Point-of-Care Assay
    Article Snippet: RT-LAMP Assay All RT-LAMP assays were assembled in 25 µL reactions containing 1× isothermal buffer supplemented with 1.4 mM deoxyribonucleotides (dNTPs), 0.4 M betaine, 6 mM additional MgSO4 , 16 units of Bst 2.0 DNA polymerase (NEB), and 7.5 units of warmstart RTx reverse transcriptase (NEB). .. Primer-containing multiplex LAMP reactions received 0.6 µM of each of the four FIP and BIP primers (NS5 BIP primer was added at 1.2 µM concentration), 0.3 µM of each of the four loop primers, 0.3 µM of the capsid stem primer, and 0.15 µM each of the four F3 and B3 primers.

    Article Title: Simultaneous Detection of Different Zika Virus Lineages via Molecular Computation in a Point-of-Care Assay
    Article Snippet: All RT-LAMP assays were assembled in 25 µL reactions containing 1× isothermal buffer supplemented with 1.4 mM deoxyribonucleotides (dNTPs), 0.4 M betaine, 6 mM additional MgSO4 , 16 units of Bst 2.0 DNA polymerase (NEB), and 7.5 units of warmstart RTx reverse transcriptase (NEB). .. Primer-containing multiplex LAMP reactions received 0.6 µM of each of the four FIP and BIP primers ( NS5 BIP primer was added at 1.2 µM concentration), 0.3 µM of each of the four loop primers, 0.3 µM of the capsid stem primer, and 0.15 µM each of the four F3 and B3 primers.

    Amplification:

    Article Title: Ultrasensitive loop mediated isothermal amplification (US-LAMP) to detect malaria for elimination
    Article Snippet: .. Ultrasensitive loop mediated amplification (US-LAMP) assay conditions Bst 2.0 WarmStart® DNA polymerase was combined with WarmStart® reverse transcriptase in 1X Isothermal Amplification Buffer (New England Biolabs, Whitby, ON) to perform the US-LAMP assay. .. In a 25-µL LAMP reaction mixture, 1.6 µM F1P and B1P, 0.8 µM LPF and LPB, 0.2 µM F3 and B3 primer concentrations, 8 mM MgSO4 , 1.4 mM dNTPs, 0.8 M Betaine (Sigma-Aldrich, Oakville, ON, Canada), 8 unit of Bst 2.0 WarmStart® DNA Polymerase and 7.5 unit of Warm Start® reverse transcriptase were used.

    Agarose Gel Electrophoresis:

    Article Title: Point of sampling detection of Zika virus within a multiplexed kit capable of detecting dengue and chikungunya
    Article Snippet: RT-LAMP procedure Reaction mixtures (50 μL total volume) contained a 10X primer set (5 μL, 16 μM FIP and BIP, 2 μM F3 and B3, 5 μM LF (or LB for chikungunya), 2 μM LB (or LF for chikungunya), 4 μM LF quencher probe, and 3 μM LB-fluorescent probe (or LF probe for chikungunya)), deoxynucleoside triphosphates (dNTPs, 1.4 mM of each), Tris-HCl buffer (20 mM, pH 8.8), KCl (50 mM), (NH4 )2 SO4 (10 mM,) MgSO4 (8 mM), Tween® 20 (0.1%), DTT (1 mM), Bst 2.0 WarmStart® DNA Polymerase (16 U, NEB, Ipswich, MA), WarmStart® RTx Reverse Transcriptase (15 U, NEB, Ipswich, MA), and RNaseOUT™ recombinant ribonuclease inhibitor (80 U, Thermo Fisher Scientific, Waltham, MA). .. Samples were incubated at 65 °C for 45 min, then analyzed by agarose gel electrophoresis (2.5%) in 1X TBE buffer, followed by ethidium bromide staining, using an appropriate DNA size marker (50 bp ladder; Promega, Madison, WI).

    Fluorescence:

    Article Title: Development and evaluation of a rapid molecular diagnostic test for Zika virus infection by reverse transcription loop-mediated isothermal amplification
    Article Snippet: RT-LAMP RT-LAMP was performed with Isothermal Master Mix reagent (Optigene, West Sussex, UK) using the Genelyzer FIII real-time fluorescence detection platform (TOSHIBA Medical Systems, Otawara, Japan). .. The reaction mixture (total volume, 25 µL) contained 15 µL of Isothermal Master Mix; 1 µL of WarmStart RTx reverse transcriptase (1 U; New England BioLabs, Ipswich, MA, USA); 4 µL of the LAMP primer mix consisting of 5 pmol F3 and B3, 20 pmol FIP and BIP, 10 pmol LF and LB; and 5 µL of RNA sample (template).

    Article Title: Ultrasensitive loop mediated isothermal amplification (US-LAMP) to detect malaria for elimination
    Article Snippet: Ultrasensitive loop mediated amplification (US-LAMP) assay conditions Bst 2.0 WarmStart® DNA polymerase was combined with WarmStart® reverse transcriptase in 1X Isothermal Amplification Buffer (New England Biolabs, Whitby, ON) to perform the US-LAMP assay. .. Amplification was measured based on increased relative fluorescence units (RFU) per minute in the CFX-96 Real-Time PCR detection system (Bio-Rad, Mississauga, ON).

    Polymerase Chain Reaction:

    Article Title: Bactericidal Activity of Lactic Acid against Clinical, Carbapenem-Hydrolyzing, Multi-Drug-Resistant Klebsiella pneumoniae Planktonic and Biofilm-Forming Cells
    Article Snippet: .. First strand cDNA synthesis was carried out using 500 ng total RNA, 500 µM of each deoxynucleoside triphosphate (dNTP; New England Biolabs, Ipswich, MA, USA), 200 ng random hexamers (Thermo Fisher Scientific, Waltham, MA, USA), and 3.75 U WarmStart® RTx Reverse Transcriptase (New England Biolabs), followed by semi-quantitative reverse transcriptase PCR (RT-PCR), as described previously and DNA Polymerase I gene expression was used as an internal control [ , ]. .. Time Course LA MIC Determination Assays on Planktonic Cells In-vitro broth dilution assays were initially carried out in duplicate using ~108 JNM11.C4 planktonic cells grown in Muller-Hinton broth (MHB; Himedia Labs).

    RT Lamp Assay:

    Article Title: Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva
    Article Snippet: Paragraph title: RT-LAMP assay ... RT-LAMP reaction was carried out in a final volume of 25 μl including the OptiGene Master mix ISO-004 with 7.5 units of WarmStart reverse transcriptase (New England BioLabs, M0380L) and 3 pairs of primers targeting a unique conserved sequence in the ZIKV capsid ( ).

    Article Title: Simultaneous Detection of Different Zika Virus Lineages via Molecular Computation in a Point-of-Care Assay
    Article Snippet: .. RT-LAMP Assay All RT-LAMP assays were assembled in 25 µL reactions containing 1× isothermal buffer supplemented with 1.4 mM deoxyribonucleotides (dNTPs), 0.4 M betaine, 6 mM additional MgSO4 , 16 units of Bst 2.0 DNA polymerase (NEB), and 7.5 units of warmstart RTx reverse transcriptase (NEB). ..

    Article Title: Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva
    Article Snippet: .. RT-LAMP assay RT-LAMP reaction was carried out in a final volume of 25 μl including the OptiGene Master mix ISO-004 with 7.5 units of WarmStart reverse transcriptase (New England BioLabs, M0380L) and 3 pairs of primers targeting a unique conserved sequence in the ZIKV capsid ( ). ..

    Article Title: Rapid and Specific Detection of All Known Nipah virus Strains’ Sequences With Reverse Transcription-Loop-Mediated Isothermal Amplification
    Article Snippet: .. RT-LAMP Assay The RT-LAMP reaction was performed in a 25.0 μl volume, containing 0.5 mM betaine (5 mM stock, Sigma-Aldrich Inc., St Louis, MO, United States), 1.4 mM each dNTP (10.0 mM stock, TaKaRa Bio Inc., Clontech Laboratories, Inc., Dalian, China), 2.5 μl of 10× ThermoPol Reaction Buffer [(1 × ThermoPol Reaction Buffer contains 20.0 mM Tris–HCl, pH 8.8; 10.0 mM KCl; 2.0 mM MgSO4 ; 10.0 mM (NH4 )2 SO4 ; 0.1% Triton X-100) New England Biolabs Inc.], 6.0 mM MgSO4 (New England Biolabs Inc.), 32 pM each of forward inner primer (FIP) and backward inner primer (BIP), 15 pM each of forward loop (LF) and backward loop (LB) primers (or replaced with an equal volume of nuclease-free water), 4 pM each of forward outer (F3) and backward outer (B3) primers, 8.0 U of Bst DNA polymerase large fragment (New England Biolabs Inc.), 7.5 U of WarmStart RTx reverse transcriptase (New England Biolabs Inc.; only added to the reaction system when the template was RNA), and 2.0 μl of pseudovirus RNA (≥50 pg/μl). .. The RNA from the cell extract transfected with pHAGE-CMV-MCS-IZsGreen vector and nuclease-free water were used as the negative controls, and the plasmids containing the three N genes were used as the positive controls (mixed without 7.5 U of WarmStart RTx reverse transcriptase).

    Article Title: Simultaneous Detection of Different Zika Virus Lineages via Molecular Computation in a Point-of-Care Assay
    Article Snippet: Paragraph title: 2.7. RT-LAMP Assay ... All RT-LAMP assays were assembled in 25 µL reactions containing 1× isothermal buffer supplemented with 1.4 mM deoxyribonucleotides (dNTPs), 0.4 M betaine, 6 mM additional MgSO4 , 16 units of Bst 2.0 DNA polymerase (NEB), and 7.5 units of warmstart RTx reverse transcriptase (NEB).

    Isolation:

    Article Title: Bactericidal Activity of Lactic Acid against Clinical, Carbapenem-Hydrolyzing, Multi-Drug-Resistant Klebsiella pneumoniae Planktonic and Biofilm-Forming Cells
    Article Snippet: For this experiment, each isolate was grown in five independent wells and the cells were pooled together for RNA isolation. .. First strand cDNA synthesis was carried out using 500 ng total RNA, 500 µM of each deoxynucleoside triphosphate (dNTP; New England Biolabs, Ipswich, MA, USA), 200 ng random hexamers (Thermo Fisher Scientific, Waltham, MA, USA), and 3.75 U WarmStart® RTx Reverse Transcriptase (New England Biolabs), followed by semi-quantitative reverse transcriptase PCR (RT-PCR), as described previously and DNA Polymerase I gene expression was used as an internal control [ , ].

    Marker:

    Article Title: Point of sampling detection of Zika virus within a multiplexed kit capable of detecting dengue and chikungunya
    Article Snippet: RT-LAMP procedure Reaction mixtures (50 μL total volume) contained a 10X primer set (5 μL, 16 μM FIP and BIP, 2 μM F3 and B3, 5 μM LF (or LB for chikungunya), 2 μM LB (or LF for chikungunya), 4 μM LF quencher probe, and 3 μM LB-fluorescent probe (or LF probe for chikungunya)), deoxynucleoside triphosphates (dNTPs, 1.4 mM of each), Tris-HCl buffer (20 mM, pH 8.8), KCl (50 mM), (NH4 )2 SO4 (10 mM,) MgSO4 (8 mM), Tween® 20 (0.1%), DTT (1 mM), Bst 2.0 WarmStart® DNA Polymerase (16 U, NEB, Ipswich, MA), WarmStart® RTx Reverse Transcriptase (15 U, NEB, Ipswich, MA), and RNaseOUT™ recombinant ribonuclease inhibitor (80 U, Thermo Fisher Scientific, Waltham, MA). .. Samples were incubated at 65 °C for 45 min, then analyzed by agarose gel electrophoresis (2.5%) in 1X TBE buffer, followed by ethidium bromide staining, using an appropriate DNA size marker (50 bp ladder; Promega, Madison, WI).

    Article Title: Bactericidal Activity of Lactic Acid against Clinical, Carbapenem-Hydrolyzing, Multi-Drug-Resistant Klebsiella pneumoniae Planktonic and Biofilm-Forming Cells
    Article Snippet: To further validate the presence of biofilm-forming cells, mrkA gene expression was used as a marker. .. First strand cDNA synthesis was carried out using 500 ng total RNA, 500 µM of each deoxynucleoside triphosphate (dNTP; New England Biolabs, Ipswich, MA, USA), 200 ng random hexamers (Thermo Fisher Scientific, Waltham, MA, USA), and 3.75 U WarmStart® RTx Reverse Transcriptase (New England Biolabs), followed by semi-quantitative reverse transcriptase PCR (RT-PCR), as described previously and DNA Polymerase I gene expression was used as an internal control [ , ].

    Sequencing:

    Article Title: Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva
    Article Snippet: .. RT-LAMP reaction was carried out in a final volume of 25 μl including the OptiGene Master mix ISO-004 with 7.5 units of WarmStart reverse transcriptase (New England BioLabs, M0380L) and 3 pairs of primers targeting a unique conserved sequence in the ZIKV capsid ( ). ..

    Article Title: Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva
    Article Snippet: .. RT-LAMP assay RT-LAMP reaction was carried out in a final volume of 25 μl including the OptiGene Master mix ISO-004 with 7.5 units of WarmStart reverse transcriptase (New England BioLabs, M0380L) and 3 pairs of primers targeting a unique conserved sequence in the ZIKV capsid ( ). ..

    Transfection:

    Article Title: Rapid and Specific Detection of All Known Nipah virus Strains’ Sequences With Reverse Transcription-Loop-Mediated Isothermal Amplification
    Article Snippet: RT-LAMP Assay The RT-LAMP reaction was performed in a 25.0 μl volume, containing 0.5 mM betaine (5 mM stock, Sigma-Aldrich Inc., St Louis, MO, United States), 1.4 mM each dNTP (10.0 mM stock, TaKaRa Bio Inc., Clontech Laboratories, Inc., Dalian, China), 2.5 μl of 10× ThermoPol Reaction Buffer [(1 × ThermoPol Reaction Buffer contains 20.0 mM Tris–HCl, pH 8.8; 10.0 mM KCl; 2.0 mM MgSO4 ; 10.0 mM (NH4 )2 SO4 ; 0.1% Triton X-100) New England Biolabs Inc.], 6.0 mM MgSO4 (New England Biolabs Inc.), 32 pM each of forward inner primer (FIP) and backward inner primer (BIP), 15 pM each of forward loop (LF) and backward loop (LB) primers (or replaced with an equal volume of nuclease-free water), 4 pM each of forward outer (F3) and backward outer (B3) primers, 8.0 U of Bst DNA polymerase large fragment (New England Biolabs Inc.), 7.5 U of WarmStart RTx reverse transcriptase (New England Biolabs Inc.; only added to the reaction system when the template was RNA), and 2.0 μl of pseudovirus RNA (≥50 pg/μl). .. The RNA from the cell extract transfected with pHAGE-CMV-MCS-IZsGreen vector and nuclease-free water were used as the negative controls, and the plasmids containing the three N genes were used as the positive controls (mixed without 7.5 U of WarmStart RTx reverse transcriptase).

    SYBR Green Assay:

    Article Title: Ultrasensitive loop mediated isothermal amplification (US-LAMP) to detect malaria for elimination
    Article Snippet: Ultrasensitive loop mediated amplification (US-LAMP) assay conditions Bst 2.0 WarmStart® DNA polymerase was combined with WarmStart® reverse transcriptase in 1X Isothermal Amplification Buffer (New England Biolabs, Whitby, ON) to perform the US-LAMP assay. .. The assay was optimized with pre-addition of 0.5 µL of 50X SYBR green (Invitrogen, Burlington, ON) in the reaction mixture.

    Concentration Assay:

    Article Title: Simultaneous Detection of Different Zika Virus Lineages via Molecular Computation in a Point-of-Care Assay
    Article Snippet: RT-LAMP Assay All RT-LAMP assays were assembled in 25 µL reactions containing 1× isothermal buffer supplemented with 1.4 mM deoxyribonucleotides (dNTPs), 0.4 M betaine, 6 mM additional MgSO4 , 16 units of Bst 2.0 DNA polymerase (NEB), and 7.5 units of warmstart RTx reverse transcriptase (NEB). .. Primer-containing multiplex LAMP reactions received 0.6 µM of each of the four FIP and BIP primers (NS5 BIP primer was added at 1.2 µM concentration), 0.3 µM of each of the four loop primers, 0.3 µM of the capsid stem primer, and 0.15 µM each of the four F3 and B3 primers.

    Article Title: Simultaneous Detection of Different Zika Virus Lineages via Molecular Computation in a Point-of-Care Assay
    Article Snippet: All RT-LAMP assays were assembled in 25 µL reactions containing 1× isothermal buffer supplemented with 1.4 mM deoxyribonucleotides (dNTPs), 0.4 M betaine, 6 mM additional MgSO4 , 16 units of Bst 2.0 DNA polymerase (NEB), and 7.5 units of warmstart RTx reverse transcriptase (NEB). .. Primer-containing multiplex LAMP reactions received 0.6 µM of each of the four FIP and BIP primers ( NS5 BIP primer was added at 1.2 µM concentration), 0.3 µM of each of the four loop primers, 0.3 µM of the capsid stem primer, and 0.15 µM each of the four F3 and B3 primers.

    Incubation:

    Article Title: Point of sampling detection of Zika virus within a multiplexed kit capable of detecting dengue and chikungunya
    Article Snippet: RT-LAMP procedure Reaction mixtures (50 μL total volume) contained a 10X primer set (5 μL, 16 μM FIP and BIP, 2 μM F3 and B3, 5 μM LF (or LB for chikungunya), 2 μM LB (or LF for chikungunya), 4 μM LF quencher probe, and 3 μM LB-fluorescent probe (or LF probe for chikungunya)), deoxynucleoside triphosphates (dNTPs, 1.4 mM of each), Tris-HCl buffer (20 mM, pH 8.8), KCl (50 mM), (NH4 )2 SO4 (10 mM,) MgSO4 (8 mM), Tween® 20 (0.1%), DTT (1 mM), Bst 2.0 WarmStart® DNA Polymerase (16 U, NEB, Ipswich, MA), WarmStart® RTx Reverse Transcriptase (15 U, NEB, Ipswich, MA), and RNaseOUT™ recombinant ribonuclease inhibitor (80 U, Thermo Fisher Scientific, Waltham, MA). .. Samples were incubated at 65 °C for 45 min, then analyzed by agarose gel electrophoresis (2.5%) in 1X TBE buffer, followed by ethidium bromide staining, using an appropriate DNA size marker (50 bp ladder; Promega, Madison, WI).

    Article Title: Rapid and Specific Detection of All Known Nipah virus Strains’ Sequences With Reverse Transcription-Loop-Mediated Isothermal Amplification
    Article Snippet: RT-LAMP Assay The RT-LAMP reaction was performed in a 25.0 μl volume, containing 0.5 mM betaine (5 mM stock, Sigma-Aldrich Inc., St Louis, MO, United States), 1.4 mM each dNTP (10.0 mM stock, TaKaRa Bio Inc., Clontech Laboratories, Inc., Dalian, China), 2.5 μl of 10× ThermoPol Reaction Buffer [(1 × ThermoPol Reaction Buffer contains 20.0 mM Tris–HCl, pH 8.8; 10.0 mM KCl; 2.0 mM MgSO4 ; 10.0 mM (NH4 )2 SO4 ; 0.1% Triton X-100) New England Biolabs Inc.], 6.0 mM MgSO4 (New England Biolabs Inc.), 32 pM each of forward inner primer (FIP) and backward inner primer (BIP), 15 pM each of forward loop (LF) and backward loop (LB) primers (or replaced with an equal volume of nuclease-free water), 4 pM each of forward outer (F3) and backward outer (B3) primers, 8.0 U of Bst DNA polymerase large fragment (New England Biolabs Inc.), 7.5 U of WarmStart RTx reverse transcriptase (New England Biolabs Inc.; only added to the reaction system when the template was RNA), and 2.0 μl of pseudovirus RNA (≥50 pg/μl). .. The mixture was covered with a grain of sealant (low-melting-point wax, melting point: 48.0–50.0°C), and incubated for 60 min in a Realtime Turbidimeter LA-320C (Eiken Chemical Co., Ltd., Tochigi, Japan) to collect the turbidity data at 61–65°C, followed by incubation at 75°C for 5 min to inactivate the enzymes.

    Expressing:

    Article Title: Bactericidal Activity of Lactic Acid against Clinical, Carbapenem-Hydrolyzing, Multi-Drug-Resistant Klebsiella pneumoniae Planktonic and Biofilm-Forming Cells
    Article Snippet: .. First strand cDNA synthesis was carried out using 500 ng total RNA, 500 µM of each deoxynucleoside triphosphate (dNTP; New England Biolabs, Ipswich, MA, USA), 200 ng random hexamers (Thermo Fisher Scientific, Waltham, MA, USA), and 3.75 U WarmStart® RTx Reverse Transcriptase (New England Biolabs), followed by semi-quantitative reverse transcriptase PCR (RT-PCR), as described previously and DNA Polymerase I gene expression was used as an internal control [ , ]. .. Time Course LA MIC Determination Assays on Planktonic Cells In-vitro broth dilution assays were initially carried out in duplicate using ~108 JNM11.C4 planktonic cells grown in Muller-Hinton broth (MHB; Himedia Labs).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Bactericidal Activity of Lactic Acid against Clinical, Carbapenem-Hydrolyzing, Multi-Drug-Resistant Klebsiella pneumoniae Planktonic and Biofilm-Forming Cells
    Article Snippet: .. First strand cDNA synthesis was carried out using 500 ng total RNA, 500 µM of each deoxynucleoside triphosphate (dNTP; New England Biolabs, Ipswich, MA, USA), 200 ng random hexamers (Thermo Fisher Scientific, Waltham, MA, USA), and 3.75 U WarmStart® RTx Reverse Transcriptase (New England Biolabs), followed by semi-quantitative reverse transcriptase PCR (RT-PCR), as described previously and DNA Polymerase I gene expression was used as an internal control [ , ]. .. Time Course LA MIC Determination Assays on Planktonic Cells In-vitro broth dilution assays were initially carried out in duplicate using ~108 JNM11.C4 planktonic cells grown in Muller-Hinton broth (MHB; Himedia Labs).

    Staining:

    Article Title: Point of sampling detection of Zika virus within a multiplexed kit capable of detecting dengue and chikungunya
    Article Snippet: RT-LAMP procedure Reaction mixtures (50 μL total volume) contained a 10X primer set (5 μL, 16 μM FIP and BIP, 2 μM F3 and B3, 5 μM LF (or LB for chikungunya), 2 μM LB (or LF for chikungunya), 4 μM LF quencher probe, and 3 μM LB-fluorescent probe (or LF probe for chikungunya)), deoxynucleoside triphosphates (dNTPs, 1.4 mM of each), Tris-HCl buffer (20 mM, pH 8.8), KCl (50 mM), (NH4 )2 SO4 (10 mM,) MgSO4 (8 mM), Tween® 20 (0.1%), DTT (1 mM), Bst 2.0 WarmStart® DNA Polymerase (16 U, NEB, Ipswich, MA), WarmStart® RTx Reverse Transcriptase (15 U, NEB, Ipswich, MA), and RNaseOUT™ recombinant ribonuclease inhibitor (80 U, Thermo Fisher Scientific, Waltham, MA). .. Samples were incubated at 65 °C for 45 min, then analyzed by agarose gel electrophoresis (2.5%) in 1X TBE buffer, followed by ethidium bromide staining, using an appropriate DNA size marker (50 bp ladder; Promega, Madison, WI).

    Real-time Polymerase Chain Reaction:

    Article Title: Ultrasensitive loop mediated isothermal amplification (US-LAMP) to detect malaria for elimination
    Article Snippet: Ultrasensitive loop mediated amplification (US-LAMP) assay conditions Bst 2.0 WarmStart® DNA polymerase was combined with WarmStart® reverse transcriptase in 1X Isothermal Amplification Buffer (New England Biolabs, Whitby, ON) to perform the US-LAMP assay. .. Amplification was measured based on increased relative fluorescence units (RFU) per minute in the CFX-96 Real-Time PCR detection system (Bio-Rad, Mississauga, ON).

    Recombinant:

    Article Title: Point of sampling detection of Zika virus within a multiplexed kit capable of detecting dengue and chikungunya
    Article Snippet: .. RT-LAMP procedure Reaction mixtures (50 μL total volume) contained a 10X primer set (5 μL, 16 μM FIP and BIP, 2 μM F3 and B3, 5 μM LF (or LB for chikungunya), 2 μM LB (or LF for chikungunya), 4 μM LF quencher probe, and 3 μM LB-fluorescent probe (or LF probe for chikungunya)), deoxynucleoside triphosphates (dNTPs, 1.4 mM of each), Tris-HCl buffer (20 mM, pH 8.8), KCl (50 mM), (NH4 )2 SO4 (10 mM,) MgSO4 (8 mM), Tween® 20 (0.1%), DTT (1 mM), Bst 2.0 WarmStart® DNA Polymerase (16 U, NEB, Ipswich, MA), WarmStart® RTx Reverse Transcriptase (15 U, NEB, Ipswich, MA), and RNaseOUT™ recombinant ribonuclease inhibitor (80 U, Thermo Fisher Scientific, Waltham, MA). ..

    Plasmid Preparation:

    Article Title: Rapid and Specific Detection of All Known Nipah virus Strains’ Sequences With Reverse Transcription-Loop-Mediated Isothermal Amplification
    Article Snippet: RT-LAMP Assay The RT-LAMP reaction was performed in a 25.0 μl volume, containing 0.5 mM betaine (5 mM stock, Sigma-Aldrich Inc., St Louis, MO, United States), 1.4 mM each dNTP (10.0 mM stock, TaKaRa Bio Inc., Clontech Laboratories, Inc., Dalian, China), 2.5 μl of 10× ThermoPol Reaction Buffer [(1 × ThermoPol Reaction Buffer contains 20.0 mM Tris–HCl, pH 8.8; 10.0 mM KCl; 2.0 mM MgSO4 ; 10.0 mM (NH4 )2 SO4 ; 0.1% Triton X-100) New England Biolabs Inc.], 6.0 mM MgSO4 (New England Biolabs Inc.), 32 pM each of forward inner primer (FIP) and backward inner primer (BIP), 15 pM each of forward loop (LF) and backward loop (LB) primers (or replaced with an equal volume of nuclease-free water), 4 pM each of forward outer (F3) and backward outer (B3) primers, 8.0 U of Bst DNA polymerase large fragment (New England Biolabs Inc.), 7.5 U of WarmStart RTx reverse transcriptase (New England Biolabs Inc.; only added to the reaction system when the template was RNA), and 2.0 μl of pseudovirus RNA (≥50 pg/μl). .. The RNA from the cell extract transfected with pHAGE-CMV-MCS-IZsGreen vector and nuclease-free water were used as the negative controls, and the plasmids containing the three N genes were used as the positive controls (mixed without 7.5 U of WarmStart RTx reverse transcriptase).

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  • 95
    New England Biolabs warmstart reverse transcriptase
    Warmstart Reverse Transcriptase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/warmstart reverse transcriptase/product/New England Biolabs
    Average 95 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    warmstart reverse transcriptase - by Bioz Stars, 2020-02
    95/100 stars
      Buy from Supplier

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