tween 20  (New England Biolabs)


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    Structured Review

    New England Biolabs tween 20
    Tween 20, 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/tween 20/product/New England Biolabs
    Average 99 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    tween 20 - by Bioz Stars, 2020-02
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    Related Articles

    Amplification:

    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: Rapid and Real-Time Detection of Chikungunya Virus by Reverse Transcription Loop-Mediated Isothermal Amplification Assay ▿
    Article Snippet: .. The RT-LAMP reaction was carried out in a total 25-μl reaction volume using the Loopamp RNA amplification kit (Eiken Chemical Co. Ltd., Japan) containing 50 pmol each of the primers FIP and BIP, 5 pmol each of the outer primers F3 and B3, 25 pmol each of loop primers F and B, 1.4 mM deoxynucleoside triphosphates, 0.8 M betaine, 0.1% Tween 20, 10 mM (NH4 )2 SO4 , 8 mM MgSO4 , 10 mM KCl, 20 mM Tris-HCl (pH 8.8), 8 units of Bst DNA polymerase (New England Biolabs), 0.625 units of AMV reverse transcriptase (Invitrogen), and 2 μl of RNA template. .. The real-time monitoring of the RT-LAMP assay was accomplished by incubating the reaction mixture at 63°C for 60 min in a Loopamp real-time turbidimeter (LA-200; Teramecs, Japan).

    Article Title: CUL1 regulates TOC1 protein stability in the Arabidopsis circadian clock
    Article Snippet: This cDNA was diluted 1:4 to produce a working stock, and 2 µl of the working stock was used as template for PCR amplification using a iCycler iQ Real-Time PCR Detection System (Bio-Rad, ), to determine the expression of TOC1, LHY , and CCA1 using the conditions, primers and probes described previously ( ). .. For this primer set and the corresponding IPP2 controls, PCR conditions were 95°C for 1 min, followed by 40 cycles of 10 sec at 95°C and 30 sec at 60°C in a buffer consisting of 1 × Ex Taq buffer (Takara Bio USA, ), 1 × EvaGreen (Biotium, ), 0.1% v/v Tween-20, 5% v/v DMSO, 50 µg ml−1 BSA (New England Biolabs, ), 0.2 mM dTNPs, 0.3 µ m primers and 0.5 U Taq polymerase (New England Biolabs).

    Article Title: Dual-Priming Isothermal Amplification (DAMP) for Highly Sensitive and Specific Molecular Detection with Ultralow Nonspecific Signals
    Article Snippet: .. Deoxynucleotide (dNTP) solution mix (10 mM of each), Mg2 SO4 (100 mM), Bacillus stearothermophilus (Bst) 2.0 WarmStart® DNA polymerase (8000 U/mL), WarmStart® RTx Reverse Transcriptase (15,000 units/mL), extreme thermostable single-stranded DNA binding protein (ET SSB, 500 μg/mL), 10 × Isothermal Amplification Buffer (200 mM Tris-HCl, 500 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 , 1.0% Tween® 20 and pH 8.8 at 25°C), and 10 ThermoPol Reaction Buffer (200 mM Tris-HCl, 100 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 and 1% Triton X-100, pH 8.8 at 25°C) were all purchased from New England BioLabs (Ipswich, MA). .. RNeasy Mini Kit for RNA extraction was purchased from QIAGEN (Frederick, MD).

    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: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes. .. Amplification products were diluted to 10X SSC and run through a dot blot apparatus onto a Hybond N+ membrande using a BioRad dot blot vacuum manifold.

    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. ..

    Real-time Polymerase Chain Reaction:

    Article Title: TPP Combined with DGUC as an Economic and Universal Process for Large-Scale Purification of AAV Vectors
    Article Snippet: .. Briefly, AAV samples were diluted to a suitable range in quantitative real-time PCR dilution buffer (10 mM Tris-HCl [pH 8.0], 1 mM EDTA, 10 μg/mL yeast tRNA, 0.01% Tween 80) and 10 μL of the diluted AAV samples was mixed with 39 μL of DNaseI digestion buffer (10mM Tris-HCl [pH 7.6], 2.5 mM MgCl2 , 0.5 mM CaCl2 ) and 1 μL (2 U) of DNaseI (New England Biolabs). .. A National Reference Standard Stock (RSS) rAAV2-RSS (ATCC) was included in all quantitative real-time PCR assays as an internal control.

    Article Title: CUL1 regulates TOC1 protein stability in the Arabidopsis circadian clock
    Article Snippet: Paragraph title: Expression analysis by real-time PCR ... For this primer set and the corresponding IPP2 controls, PCR conditions were 95°C for 1 min, followed by 40 cycles of 10 sec at 95°C and 30 sec at 60°C in a buffer consisting of 1 × Ex Taq buffer (Takara Bio USA, ), 1 × EvaGreen (Biotium, ), 0.1% v/v Tween-20, 5% v/v DMSO, 50 µg ml−1 BSA (New England Biolabs, ), 0.2 mM dTNPs, 0.3 µ m primers and 0.5 U Taq polymerase (New England Biolabs).

    Incubation:

    Article Title: A sharp Pif1-dependent threshold separates DNA double-strand breaks from critically short telomeres
    Article Snippet: 1 µL of tailing mix (0.05 µL Terminal Transferase (NEB, cat. no. M0315), 0.1 µL 10x NEBuffer 4, 0.1 µL 10 mM dCTP, 0.75 µL dH2 O) was added and incubated for 30 min at 37°C, 10 min at 65°C, and 5 min at 96°C. .. The PCR mix consisted of 4 µL 10x PCR buffer (670 mM Tris-HCl pH 8.8, 160 mM (NH4 )2 SO4 , 50% glycerol, 0.1% Tween-20), 0.32 µL 25 mM dNTP mix, 0.3 µL 100 µM telomere-specific primer (V-R: 5’-GTGAGCGGATAACAATTTCACACAGTCTAGATGTCCGAATTGATCCCAGAGTAG-3’ or VI-R: 5’-ACGTGTGCGTACGCCATATCAATATGC-3’), 0.3 µL 100 µM G18 primer (5’-CGGGATCCG18 -3’), 0.5 µL Q5 High-Fidelity DNA Polymerase (NEB, cat. no. M0491), 24.68 µL dH2 O.

    Article Title: Rapid and Precise Engineering of the Caenorhabditis elegans Genome with Lethal Mutation Co-Conversion and Inactivation of NHEJ Repair
    Article Snippet: .. Genotyping PCRs and restriction digestion Single F1 animals were picked into 10 µl of single-worm lysis buffer [10 mM Tris (pH 8.3), 50 mM KCl, 2.5 mM MgCl2 , 0.45% IGEPAL, 0.45% Tween-20] containing 1 mg/ml proteinase K. Tubes were incubated on dry ice for 15 min, 62° for 1 hr, and then heated to 95° for 20 min to inactivate the proteinase K. Genotyping PCRs were performed using Phusion polymerase (NEB, no. M0530S) and High Fidelity buffer; 1/10 volume of lysate was used as template in the PCR. .. For identification of knock-in events in the F1 , 30-µl PCRs were performed.

    Article Title: An Olfactory Sensory Neuron Line, Odora, Properly Targets Olfactory Proteins and Responds to Odorants
    Article Snippet: .. The membrane was blocked in 5% (w/v) nonfat milk in 200 m m NaCl, 0.1% Tween 20, 50 m m Tris, pH 7.4 for 1 hr, incubated in primary antibody for 2 hr and peroxidase-conjugated secondary antibody (New England Biolabs, Beverly, MA) for 1 hr, all at ambient temperature. .. After washing, the membrane was incubated in a chemiluminescent substrate (New England Nuclear, Boston, MA) for 1 min and exposed to x-ray film.

    Article Title: Mammalian Homolog of Drosophila retinal degeneration B Rescues the Mutant Fly Phenotype
    Article Snippet: .. After several washes with 1× PBS and 0.1% Tween-20, the membrane was incubated with peroxidase-conjugated anti-rabbit secondary antibody (New England Biolabs, 1:1000), and the signal was detected using an ECL kit as described by the manufacturer (Amersham). ..

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: .. Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes. .. 80 ng of gDNA was incubated in the presence of Φ29 polymerase, and 20 ng of gDNA was incubated in the absence of Φ29 polymerase.

    Article Title: Constitutively bound CTCF sites maintain 3D chromatin architecture and long-range epigenetically regulated domains
    Article Snippet: This material was added 1:1 to MyOne Streptavidin C1 beads (#650-01, Invitrogen) that had been resuspended in 2× binding buffer (10 mM Tris–HCl pH 8.0, 1 mM EDTA and 2 M NaCl) and incubated for 20 min at room temperature, with rotation. .. Adapter-ligated DNA was washed 2× with 200 μl of 1× tween wash buffer (5 mM Tris–HCl pH 8.0, 0.5 mM EDTA, 1 M NaCl and 0.05% Tween (#p7949, Sigma Aldrich)), 1× with 200 μl binding buffer and 1× with 200 μl 1× NEB2 buffer (#B7002S, NEB) before beads were resuspended in 30 μl H2 O. PCR enrichment of adapter-ligated DNA was performed on DNA bound to the MyOne Streptavidin C1 beads using NEB kit (# E7370L, NEB).

    Expressing:

    Article Title: CUL1 regulates TOC1 protein stability in the Arabidopsis circadian clock
    Article Snippet: Paragraph title: Expression analysis by real-time PCR ... For this primer set and the corresponding IPP2 controls, PCR conditions were 95°C for 1 min, followed by 40 cycles of 10 sec at 95°C and 30 sec at 60°C in a buffer consisting of 1 × Ex Taq buffer (Takara Bio USA, ), 1 × EvaGreen (Biotium, ), 0.1% v/v Tween-20, 5% v/v DMSO, 50 µg ml−1 BSA (New England Biolabs, ), 0.2 mM dTNPs, 0.3 µ m primers and 0.5 U Taq polymerase (New England Biolabs).

    Knock-In:

    Article Title: Rapid and Precise Engineering of the Caenorhabditis elegans Genome with Lethal Mutation Co-Conversion and Inactivation of NHEJ Repair
    Article Snippet: Genotyping PCRs and restriction digestion Single F1 animals were picked into 10 µl of single-worm lysis buffer [10 mM Tris (pH 8.3), 50 mM KCl, 2.5 mM MgCl2 , 0.45% IGEPAL, 0.45% Tween-20] containing 1 mg/ml proteinase K. Tubes were incubated on dry ice for 15 min, 62° for 1 hr, and then heated to 95° for 20 min to inactivate the proteinase K. Genotyping PCRs were performed using Phusion polymerase (NEB, no. M0530S) and High Fidelity buffer; 1/10 volume of lysate was used as template in the PCR. .. For identification of knock-in events in the F1 , 30-µl PCRs were performed.

    Sequencing:

    Article Title: Dual-Priming Isothermal Amplification (DAMP) for Highly Sensitive and Specific Molecular Detection with Ultralow Nonspecific Signals
    Article Snippet: Deoxynucleotide (dNTP) solution mix (10 mM of each), Mg2 SO4 (100 mM), Bacillus stearothermophilus (Bst) 2.0 WarmStart® DNA polymerase (8000 U/mL), WarmStart® RTx Reverse Transcriptase (15,000 units/mL), extreme thermostable single-stranded DNA binding protein (ET SSB, 500 μg/mL), 10 × Isothermal Amplification Buffer (200 mM Tris-HCl, 500 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 , 1.0% Tween® 20 and pH 8.8 at 25°C), and 10 ThermoPol Reaction Buffer (200 mM Tris-HCl, 100 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 and 1% Triton X-100, pH 8.8 at 25°C) were all purchased from New England BioLabs (Ipswich, MA). .. Oligonucleotides (primers) and the pUCIDT (Amp) plasmid containing 300-bp HIV-1 p24 gene or 300-bp E. coli B mal B gene sequence were ordered from Integrated DNA Technologies (Coralville, IA).

    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.

    Protease Inhibitor:

    Article Title: Mammalian Homolog of Drosophila retinal degeneration B Rescues the Mutant Fly Phenotype
    Article Snippet: Protease inhibitor mix (10 μg/ml each leupeptin, antipain, chymostatin, and pepstatin; Sigma, St. Louis, MO) was then added to each sample. .. After several washes with 1× PBS and 0.1% Tween-20, the membrane was incubated with peroxidase-conjugated anti-rabbit secondary antibody (New England Biolabs, 1:1000), and the signal was detected using an ECL kit as described by the manufacturer (Amersham).

    SYBR Green Assay:

    Article Title: Dual-Priming Isothermal Amplification (DAMP) for Highly Sensitive and Specific Molecular Detection with Ultralow Nonspecific Signals
    Article Snippet: Agarose powder, 50 × TAE (Tris/Acetic Acid/EDTA) Buffer, and SsoAdvanced™ Universal SYBR® Green PCR Supermix were purchased from Bio-Rad Laboratories (Hercules, CA). .. Deoxynucleotide (dNTP) solution mix (10 mM of each), Mg2 SO4 (100 mM), Bacillus stearothermophilus (Bst) 2.0 WarmStart® DNA polymerase (8000 U/mL), WarmStart® RTx Reverse Transcriptase (15,000 units/mL), extreme thermostable single-stranded DNA binding protein (ET SSB, 500 μg/mL), 10 × Isothermal Amplification Buffer (200 mM Tris-HCl, 500 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 , 1.0% Tween® 20 and pH 8.8 at 25°C), and 10 ThermoPol Reaction Buffer (200 mM Tris-HCl, 100 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 and 1% Triton X-100, pH 8.8 at 25°C) were all purchased from New England BioLabs (Ipswich, MA).

    Generated:

    Article Title: CUL1 regulates TOC1 protein stability in the Arabidopsis circadian clock
    Article Snippet: First-strand cDNA was generated from 1 µg of total RNA using the SuperScript II first-strand synthesis for RT-PCR kit (Invitrogen, ). .. For this primer set and the corresponding IPP2 controls, PCR conditions were 95°C for 1 min, followed by 40 cycles of 10 sec at 95°C and 30 sec at 60°C in a buffer consisting of 1 × Ex Taq buffer (Takara Bio USA, ), 1 × EvaGreen (Biotium, ), 0.1% v/v Tween-20, 5% v/v DMSO, 50 µg ml−1 BSA (New England Biolabs, ), 0.2 mM dTNPs, 0.3 µ m primers and 0.5 U Taq polymerase (New England Biolabs).

    Article Title: Mammalian Homolog of Drosophila retinal degeneration B Rescues the Mutant Fly Phenotype
    Article Snippet: After several washes with 1× PBS and 0.1% Tween-20, the membrane was incubated with peroxidase-conjugated anti-rabbit secondary antibody (New England Biolabs, 1:1000), and the signal was detected using an ECL kit as described by the manufacturer (Amersham). .. The antisense and sense probes were generated from a pBluescript SK+ plasmid containing murine mrdgB sequences from nucleotides 1457–2138.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: CUL1 regulates TOC1 protein stability in the Arabidopsis circadian clock
    Article Snippet: First-strand cDNA was generated from 1 µg of total RNA using the SuperScript II first-strand synthesis for RT-PCR kit (Invitrogen, ). .. For this primer set and the corresponding IPP2 controls, PCR conditions were 95°C for 1 min, followed by 40 cycles of 10 sec at 95°C and 30 sec at 60°C in a buffer consisting of 1 × Ex Taq buffer (Takara Bio USA, ), 1 × EvaGreen (Biotium, ), 0.1% v/v Tween-20, 5% v/v DMSO, 50 µg ml−1 BSA (New England Biolabs, ), 0.2 mM dTNPs, 0.3 µ m primers and 0.5 U Taq polymerase (New England Biolabs).

    Sonication:

    Article Title: An Olfactory Sensory Neuron Line, Odora, Properly Targets Olfactory Proteins and Responds to Odorants
    Article Snippet: After collection and sonication, two volumes of sample buffer (192 m m Tris-HCl, pH 6.8, containing 9% SDS, 15% glycerol, and 2% 2-mercaptoethanol) were added. .. The membrane was blocked in 5% (w/v) nonfat milk in 200 m m NaCl, 0.1% Tween 20, 50 m m Tris, pH 7.4 for 1 hr, incubated in primary antibody for 2 hr and peroxidase-conjugated secondary antibody (New England Biolabs, Beverly, MA) for 1 hr, all at ambient temperature.

    Affinity Purification:

    Article Title: Mammalian Homolog of Drosophila retinal degeneration B Rescues the Mutant Fly Phenotype
    Article Snippet: The membrane was blocked for 1 hr (0.5% nonfat dry milk in 1× PBS buffer and 0.1% Tween 20) and incubated for 1 hr at room temperature with affinity-purified rabbit antibodies against murine MrdgB (1:200). .. After several washes with 1× PBS and 0.1% Tween-20, the membrane was incubated with peroxidase-conjugated anti-rabbit secondary antibody (New England Biolabs, 1:1000), and the signal was detected using an ECL kit as described by the manufacturer (Amersham).

    Binding Assay:

    Article Title: Dual-Priming Isothermal Amplification (DAMP) for Highly Sensitive and Specific Molecular Detection with Ultralow Nonspecific Signals
    Article Snippet: .. Deoxynucleotide (dNTP) solution mix (10 mM of each), Mg2 SO4 (100 mM), Bacillus stearothermophilus (Bst) 2.0 WarmStart® DNA polymerase (8000 U/mL), WarmStart® RTx Reverse Transcriptase (15,000 units/mL), extreme thermostable single-stranded DNA binding protein (ET SSB, 500 μg/mL), 10 × Isothermal Amplification Buffer (200 mM Tris-HCl, 500 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 , 1.0% Tween® 20 and pH 8.8 at 25°C), and 10 ThermoPol Reaction Buffer (200 mM Tris-HCl, 100 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 and 1% Triton X-100, pH 8.8 at 25°C) were all purchased from New England BioLabs (Ipswich, MA). .. RNeasy Mini Kit for RNA extraction was purchased from QIAGEN (Frederick, MD).

    Article Title: Constitutively bound CTCF sites maintain 3D chromatin architecture and long-range epigenetically regulated domains
    Article Snippet: .. Adapter-ligated DNA was washed 2× with 200 μl of 1× tween wash buffer (5 mM Tris–HCl pH 8.0, 0.5 mM EDTA, 1 M NaCl and 0.05% Tween (#p7949, Sigma Aldrich)), 1× with 200 μl binding buffer and 1× with 200 μl 1× NEB2 buffer (#B7002S, NEB) before beads were resuspended in 30 μl H2 O. PCR enrichment of adapter-ligated DNA was performed on DNA bound to the MyOne Streptavidin C1 beads using NEB kit (# E7370L, NEB). .. The PCR cycling steps were 1 cycle at 98 °C for 30 s, 10–14 cycles at 98 °C for 10 s/65 °C for 75 s, 1 cycle at 65 °C for 5 min. Clean up of PCR products was performed using 1x volume AMPureXP Beads (#A63881, Beckman Coulter) and products were eluted into 30 μl of nuclease-free H2 O.

    Hi-C:

    Article Title: Constitutively bound CTCF sites maintain 3D chromatin architecture and long-range epigenetically regulated domains
    Article Snippet: Paragraph title: Preparation of Hi-C libraries ... Adapter-ligated DNA was washed 2× with 200 μl of 1× tween wash buffer (5 mM Tris–HCl pH 8.0, 0.5 mM EDTA, 1 M NaCl and 0.05% Tween (#p7949, Sigma Aldrich)), 1× with 200 μl binding buffer and 1× with 200 μl 1× NEB2 buffer (#B7002S, NEB) before beads were resuspended in 30 μl H2 O. PCR enrichment of adapter-ligated DNA was performed on DNA bound to the MyOne Streptavidin C1 beads using NEB kit (# E7370L, NEB).

    DNA Extraction:

    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.

    RT Lamp Assay:

    Article Title: Rapid and Real-Time Detection of Chikungunya Virus by Reverse Transcription Loop-Mediated Isothermal Amplification Assay ▿
    Article Snippet: The RT-LAMP reaction was carried out in a total 25-μl reaction volume using the Loopamp RNA amplification kit (Eiken Chemical Co. Ltd., Japan) containing 50 pmol each of the primers FIP and BIP, 5 pmol each of the outer primers F3 and B3, 25 pmol each of loop primers F and B, 1.4 mM deoxynucleoside triphosphates, 0.8 M betaine, 0.1% Tween 20, 10 mM (NH4 )2 SO4 , 8 mM MgSO4 , 10 mM KCl, 20 mM Tris-HCl (pH 8.8), 8 units of Bst DNA polymerase (New England Biolabs), 0.625 units of AMV reverse transcriptase (Invitrogen), and 2 μl of RNA template. .. The real-time monitoring of the RT-LAMP assay was accomplished by incubating the reaction mixture at 63°C for 60 min in a Loopamp real-time turbidimeter (LA-200; Teramecs, Japan).

    Isolation:

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: Briefly, genomic DNA was isolated from 25-50 mg of frozen tumor tissue using the QIAGEN QiaAMP DNA Mini Kit according to the manufacturer’s instructions. .. Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes.

    Article Title: Constitutively bound CTCF sites maintain 3D chromatin architecture and long-range epigenetically regulated domains
    Article Snippet: Biotin-tagged DNA coupled with MyOne Streptavidin C1 beads was isolated using a magnetic particle concentrator. .. Adapter-ligated DNA was washed 2× with 200 μl of 1× tween wash buffer (5 mM Tris–HCl pH 8.0, 0.5 mM EDTA, 1 M NaCl and 0.05% Tween (#p7949, Sigma Aldrich)), 1× with 200 μl binding buffer and 1× with 200 μl 1× NEB2 buffer (#B7002S, NEB) before beads were resuspended in 30 μl H2 O. PCR enrichment of adapter-ligated DNA was performed on DNA bound to the MyOne Streptavidin C1 beads using NEB kit (# E7370L, NEB).

    Size-exclusion Chromatography:

    Article Title: CUL1 regulates TOC1 protein stability in the Arabidopsis circadian clock
    Article Snippet: .. For this primer set and the corresponding IPP2 controls, PCR conditions were 95°C for 1 min, followed by 40 cycles of 10 sec at 95°C and 30 sec at 60°C in a buffer consisting of 1 × Ex Taq buffer (Takara Bio USA, ), 1 × EvaGreen (Biotium, ), 0.1% v/v Tween-20, 5% v/v DMSO, 50 µg ml−1 BSA (New England Biolabs, ), 0.2 mM dTNPs, 0.3 µ m primers and 0.5 U Taq polymerase (New England Biolabs). ..

    Labeling:

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: Membranes were stripped in wash solution (0.5X SSC, 0.1% SDS) at 65°C and re-hybridized with γ-32P labeled Alu oligo probe 5′-GTAATCCCAGCACTTTGG-3′ for 16h at 37°C as a loading control. .. Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes.

    Purification:

    Article Title: Rapid and Precise Engineering of the Caenorhabditis elegans Genome with Lethal Mutation Co-Conversion and Inactivation of NHEJ Repair
    Article Snippet: Genotyping PCRs and restriction digestion Single F1 animals were picked into 10 µl of single-worm lysis buffer [10 mM Tris (pH 8.3), 50 mM KCl, 2.5 mM MgCl2 , 0.45% IGEPAL, 0.45% Tween-20] containing 1 mg/ml proteinase K. Tubes were incubated on dry ice for 15 min, 62° for 1 hr, and then heated to 95° for 20 min to inactivate the proteinase K. Genotyping PCRs were performed using Phusion polymerase (NEB, no. M0530S) and High Fidelity buffer; 1/10 volume of lysate was used as template in the PCR. .. CEL-1 was purified from four heads of nonorganic celery (Safeway) with the celery juice extracted with a BJE510XL 900W juicer (Breville).

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: .. Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes. .. 80 ng of gDNA was incubated in the presence of Φ29 polymerase, and 20 ng of gDNA was incubated in the absence of Φ29 polymerase.

    Dot Blot:

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes. .. Amplification products were diluted to 10X SSC and run through a dot blot apparatus onto a Hybond N+ membrande using a BioRad dot blot vacuum manifold.

    Polymerase Chain Reaction:

    Article Title: A sharp Pif1-dependent threshold separates DNA double-strand breaks from critically short telomeres
    Article Snippet: .. The PCR mix consisted of 4 µL 10x PCR buffer (670 mM Tris-HCl pH 8.8, 160 mM (NH4 )2 SO4 , 50% glycerol, 0.1% Tween-20), 0.32 µL 25 mM dNTP mix, 0.3 µL 100 µM telomere-specific primer (V-R: 5’-GTGAGCGGATAACAATTTCACACAGTCTAGATGTCCGAATTGATCCCAGAGTAG-3’ or VI-R: 5’-ACGTGTGCGTACGCCATATCAATATGC-3’), 0.3 µL 100 µM G18 primer (5’-CGGGATCCG18 -3’), 0.5 µL Q5 High-Fidelity DNA Polymerase (NEB, cat. no. M0491), 24.68 µL dH2 O. ..

    Article Title: Rapid and Precise Engineering of the Caenorhabditis elegans Genome with Lethal Mutation Co-Conversion and Inactivation of NHEJ Repair
    Article Snippet: .. Genotyping PCRs and restriction digestion Single F1 animals were picked into 10 µl of single-worm lysis buffer [10 mM Tris (pH 8.3), 50 mM KCl, 2.5 mM MgCl2 , 0.45% IGEPAL, 0.45% Tween-20] containing 1 mg/ml proteinase K. Tubes were incubated on dry ice for 15 min, 62° for 1 hr, and then heated to 95° for 20 min to inactivate the proteinase K. Genotyping PCRs were performed using Phusion polymerase (NEB, no. M0530S) and High Fidelity buffer; 1/10 volume of lysate was used as template in the PCR. .. For identification of knock-in events in the F1 , 30-µl PCRs were performed.

    Article Title: CUL1 regulates TOC1 protein stability in the Arabidopsis circadian clock
    Article Snippet: .. For this primer set and the corresponding IPP2 controls, PCR conditions were 95°C for 1 min, followed by 40 cycles of 10 sec at 95°C and 30 sec at 60°C in a buffer consisting of 1 × Ex Taq buffer (Takara Bio USA, ), 1 × EvaGreen (Biotium, ), 0.1% v/v Tween-20, 5% v/v DMSO, 50 µg ml−1 BSA (New England Biolabs, ), 0.2 mM dTNPs, 0.3 µ m primers and 0.5 U Taq polymerase (New England Biolabs). ..

    Article Title: Dual-Priming Isothermal Amplification (DAMP) for Highly Sensitive and Specific Molecular Detection with Ultralow Nonspecific Signals
    Article Snippet: Agarose powder, 50 × TAE (Tris/Acetic Acid/EDTA) Buffer, and SsoAdvanced™ Universal SYBR® Green PCR Supermix were purchased from Bio-Rad Laboratories (Hercules, CA). .. Deoxynucleotide (dNTP) solution mix (10 mM of each), Mg2 SO4 (100 mM), Bacillus stearothermophilus (Bst) 2.0 WarmStart® DNA polymerase (8000 U/mL), WarmStart® RTx Reverse Transcriptase (15,000 units/mL), extreme thermostable single-stranded DNA binding protein (ET SSB, 500 μg/mL), 10 × Isothermal Amplification Buffer (200 mM Tris-HCl, 500 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 , 1.0% Tween® 20 and pH 8.8 at 25°C), and 10 ThermoPol Reaction Buffer (200 mM Tris-HCl, 100 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 and 1% Triton X-100, pH 8.8 at 25°C) were all purchased from New England BioLabs (Ipswich, MA).

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: Following purification, genomic DNA was digested with AluI and MboI restriction enzymes overnight at 37°C, and then purified using a QIAGEN PCR clean-up kit according to the manufacturer’s instructions. .. Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes.

    Article Title: Constitutively bound CTCF sites maintain 3D chromatin architecture and long-range epigenetically regulated domains
    Article Snippet: .. Adapter-ligated DNA was washed 2× with 200 μl of 1× tween wash buffer (5 mM Tris–HCl pH 8.0, 0.5 mM EDTA, 1 M NaCl and 0.05% Tween (#p7949, Sigma Aldrich)), 1× with 200 μl binding buffer and 1× with 200 μl 1× NEB2 buffer (#B7002S, NEB) before beads were resuspended in 30 μl H2 O. PCR enrichment of adapter-ligated DNA was performed on DNA bound to the MyOne Streptavidin C1 beads using NEB kit (# E7370L, NEB). .. The PCR cycling steps were 1 cycle at 98 °C for 30 s, 10–14 cycles at 98 °C for 10 s/65 °C for 75 s, 1 cycle at 65 °C for 5 min. Clean up of PCR products was performed using 1x volume AMPureXP Beads (#A63881, Beckman Coulter) and products were eluted into 30 μl of nuclease-free H2 O.

    Plasmid Preparation:

    Article Title: Mammalian Homolog of Drosophila retinal degeneration B Rescues the Mutant Fly Phenotype
    Article Snippet: After several washes with 1× PBS and 0.1% Tween-20, the membrane was incubated with peroxidase-conjugated anti-rabbit secondary antibody (New England Biolabs, 1:1000), and the signal was detected using an ECL kit as described by the manufacturer (Amersham). .. The antisense and sense probes were generated from a pBluescript SK+ plasmid containing murine mrdgB sequences from nucleotides 1457–2138.

    Article Title: Dual-Priming Isothermal Amplification (DAMP) for Highly Sensitive and Specific Molecular Detection with Ultralow Nonspecific Signals
    Article Snippet: Deoxynucleotide (dNTP) solution mix (10 mM of each), Mg2 SO4 (100 mM), Bacillus stearothermophilus (Bst) 2.0 WarmStart® DNA polymerase (8000 U/mL), WarmStart® RTx Reverse Transcriptase (15,000 units/mL), extreme thermostable single-stranded DNA binding protein (ET SSB, 500 μg/mL), 10 × Isothermal Amplification Buffer (200 mM Tris-HCl, 500 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 , 1.0% Tween® 20 and pH 8.8 at 25°C), and 10 ThermoPol Reaction Buffer (200 mM Tris-HCl, 100 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 and 1% Triton X-100, pH 8.8 at 25°C) were all purchased from New England BioLabs (Ipswich, MA). .. Oligonucleotides (primers) and the pUCIDT (Amp) plasmid containing 300-bp HIV-1 p24 gene or 300-bp E. coli B mal B gene sequence were ordered from Integrated DNA Technologies (Coralville, IA).

    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.

    Electrophoresis:

    Article Title: An Olfactory Sensory Neuron Line, Odora, Properly Targets Olfactory Proteins and Responds to Odorants
    Article Snippet: Lysates were boiled and separated by electrophoresis on a denaturing 10% polyacrylamide gel, then transferred to Immobilon-P (Millipore, Bedford, MA). .. The membrane was blocked in 5% (w/v) nonfat milk in 200 m m NaCl, 0.1% Tween 20, 50 m m Tris, pH 7.4 for 1 hr, incubated in primary antibody for 2 hr and peroxidase-conjugated secondary antibody (New England Biolabs, Beverly, MA) for 1 hr, all at ambient temperature.

    RNA Extraction:

    Article Title: Dual-Priming Isothermal Amplification (DAMP) for Highly Sensitive and Specific Molecular Detection with Ultralow Nonspecific Signals
    Article Snippet: Deoxynucleotide (dNTP) solution mix (10 mM of each), Mg2 SO4 (100 mM), Bacillus stearothermophilus (Bst) 2.0 WarmStart® DNA polymerase (8000 U/mL), WarmStart® RTx Reverse Transcriptase (15,000 units/mL), extreme thermostable single-stranded DNA binding protein (ET SSB, 500 μg/mL), 10 × Isothermal Amplification Buffer (200 mM Tris-HCl, 500 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 , 1.0% Tween® 20 and pH 8.8 at 25°C), and 10 ThermoPol Reaction Buffer (200 mM Tris-HCl, 100 mM KCl, 100 mM (NH4 )2 SO4 , 20 mM MgSO4 and 1% Triton X-100, pH 8.8 at 25°C) were all purchased from New England BioLabs (Ipswich, MA). .. RNeasy Mini Kit for RNA extraction was purchased from QIAGEN (Frederick, MD).

    Selection:

    Article Title: Constitutively bound CTCF sites maintain 3D chromatin architecture and long-range epigenetically regulated domains
    Article Snippet: Following end repair a size selection was performed using 1.6× volume AMPureXP Beads (#A63881, Beckman Coulter Inc.). .. Adapter-ligated DNA was washed 2× with 200 μl of 1× tween wash buffer (5 mM Tris–HCl pH 8.0, 0.5 mM EDTA, 1 M NaCl and 0.05% Tween (#p7949, Sigma Aldrich)), 1× with 200 μl binding buffer and 1× with 200 μl 1× NEB2 buffer (#B7002S, NEB) before beads were resuspended in 30 μl H2 O. PCR enrichment of adapter-ligated DNA was performed on DNA bound to the MyOne Streptavidin C1 beads using NEB kit (# E7370L, NEB).

    C Circle Assay:

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: Paragraph title: C-circle Assay ... Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes.

    Spectrophotometry:

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: .. Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes. .. 80 ng of gDNA was incubated in the presence of Φ29 polymerase, and 20 ng of gDNA was incubated in the absence of Φ29 polymerase.

    Concentration Assay:

    Article Title: SLX4IP Antagonizes Promiscuous BLM Activity during ALT Maintenance
    Article Snippet: .. Purified, digested DNA was quantified with a Nanodrop spectrophotometer and then diluted to a concentration of 10 ng/μl. gDNA was diluted in 25 μl of 1X Φ29 buffer (NEB) containing BSA (NEB; 0.08 mg/ml), 0.1% Tween-20, 0.25 mM each of dATP, dGTP, and dTTP, then incubated in the presence or absence of 7.5 U Φ29 polymerase (NEB) at 30°C for 8 hours, then 65°C for 20 minutes. .. 80 ng of gDNA was incubated in the presence of Φ29 polymerase, and 20 ng of gDNA was incubated in the absence of Φ29 polymerase.

    Lysis:

    Article Title: Rapid and Precise Engineering of the Caenorhabditis elegans Genome with Lethal Mutation Co-Conversion and Inactivation of NHEJ Repair
    Article Snippet: .. Genotyping PCRs and restriction digestion Single F1 animals were picked into 10 µl of single-worm lysis buffer [10 mM Tris (pH 8.3), 50 mM KCl, 2.5 mM MgCl2 , 0.45% IGEPAL, 0.45% Tween-20] containing 1 mg/ml proteinase K. Tubes were incubated on dry ice for 15 min, 62° for 1 hr, and then heated to 95° for 20 min to inactivate the proteinase K. Genotyping PCRs were performed using Phusion polymerase (NEB, no. M0530S) and High Fidelity buffer; 1/10 volume of lysate was used as template in the PCR. .. For identification of knock-in events in the F1 , 30-µl PCRs were performed.

    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 bst 2 0 dna polymerase
    Sensitivity of the L . loa RF4-based LAMP assay. Dilutions of genomic L . loa <t>DNA</t> were amplified with the RF4 primer set and Bst 2.0 DNA polymerase in the absence (blue) or presence (red) of the V/DEF additive. Two ul of each dilution was added to LAMP reactions. The average threshold time, defined as the time at which the change in turbidity over time (dT/dt) reaches a value of 0.1, is plotted against the concentration of L . loa DNA (ng/ml). All reactions were performed in triplicate. Error bars represent the standard deviation at each point.
    Bst 2 0 Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    New England Biolabs pcmv t20 plasmid
    Antibody titers to HIV-1 gp160 subtype (B) following <t>pCMV-EnvABC</t> and <t>T20</t> peptide immunization. Six mice in each group were immunized with pCMV-EnvABC (20 µg) with T20 peptide (2 µg). Immunogens were given intramuscularly (im), intradermally (id) or intranasally (ina). Sera or tissue antibody levels were assayed by ELISA. Median IgG titers and ranges are shown.
    Pcmv T20 Plasmid, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs tris hcl
    Rate dependence of linear QPA on the length of AT-rich segment. (A) Targets and primer sequences; targets 1, 2 and 3 contain 34-nt, 20-nt and 5-nt AT-rich segments, respectively. (B) Typical dGTP-QPA (black) and dNTP-QPA profiles (colored) obtained using 0.5 μM primer, 10 nM target 2, 100 μM dGTP or 800 μM dNTP, 0.08 U/uL Bst 2.0 polymerase in 10 mM <t>KCl,</t> 40 mM CsCl, 2 mM MgCl 2 and 10 mM <t>Tris-HCl,</t> pH 8.7 at 62 °C. Dashed line corresponds to no-template control (NTC). (C) Temperature dependence of the rate of linear QPA using target 1 (blue), target 2 (red) and target 3 (black).
    Tris Hcl, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Sensitivity of the L . loa RF4-based LAMP assay. Dilutions of genomic L . loa DNA were amplified with the RF4 primer set and Bst 2.0 DNA polymerase in the absence (blue) or presence (red) of the V/DEF additive. Two ul of each dilution was added to LAMP reactions. The average threshold time, defined as the time at which the change in turbidity over time (dT/dt) reaches a value of 0.1, is plotted against the concentration of L . loa DNA (ng/ml). All reactions were performed in triplicate. Error bars represent the standard deviation at each point.

    Journal: PLoS ONE

    Article Title: Genome Filtering for New DNA Biomarkers of Loa loa Infection Suitable for Loop-Mediated Isothermal Amplification

    doi: 10.1371/journal.pone.0139286

    Figure Lengend Snippet: Sensitivity of the L . loa RF4-based LAMP assay. Dilutions of genomic L . loa DNA were amplified with the RF4 primer set and Bst 2.0 DNA polymerase in the absence (blue) or presence (red) of the V/DEF additive. Two ul of each dilution was added to LAMP reactions. The average threshold time, defined as the time at which the change in turbidity over time (dT/dt) reaches a value of 0.1, is plotted against the concentration of L . loa DNA (ng/ml). All reactions were performed in triplicate. Error bars represent the standard deviation at each point.

    Article Snippet: LAMP Assays LAMP reactions contained 1.6 μM each of primers FIP and BIP, 0.2 μM each of F3 and B3, 0.4 μM each of LF and FB, 1.4 mM of each dNTP, 20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 8 mM MgSO4 , 0.1% Tween-20 and 8 U of Bst 2.0 DNA Polymerase (New England Biolabs) mixed with one of several template DNAs, or 10 mM Tris-HCl (pH 8), 0.1 mM EDTA for non-template controls (NTC), in a total volume of 25 μl.

    Techniques: Lamp Assay, Amplification, Concentration Assay, Standard Deviation

    Detection of L . loa DNA in spiked blood samples. A two-fold dilution series of genomic L . loa DNA was prepared using uninfected human whole blood. NTCs only contained uninfected human whole blood. After DNA isolation, two μl of each dilution (or NTC) was used in LAMP reactions containing the V/DEF additive with Bst 2.0 DNA polymerase. For each experiment, all samples were assayed in triplicate. Average threshold times and standard deviations are plotted against ng DNA/ml of elution buffer.

    Journal: PLoS ONE

    Article Title: Genome Filtering for New DNA Biomarkers of Loa loa Infection Suitable for Loop-Mediated Isothermal Amplification

    doi: 10.1371/journal.pone.0139286

    Figure Lengend Snippet: Detection of L . loa DNA in spiked blood samples. A two-fold dilution series of genomic L . loa DNA was prepared using uninfected human whole blood. NTCs only contained uninfected human whole blood. After DNA isolation, two μl of each dilution (or NTC) was used in LAMP reactions containing the V/DEF additive with Bst 2.0 DNA polymerase. For each experiment, all samples were assayed in triplicate. Average threshold times and standard deviations are plotted against ng DNA/ml of elution buffer.

    Article Snippet: LAMP Assays LAMP reactions contained 1.6 μM each of primers FIP and BIP, 0.2 μM each of F3 and B3, 0.4 μM each of LF and FB, 1.4 mM of each dNTP, 20 mM Tris-HCl (pH 8.8), 50 mM KCl, 10 mM (NH4 )2 SO4 , 8 mM MgSO4 , 0.1% Tween-20 and 8 U of Bst 2.0 DNA Polymerase (New England Biolabs) mixed with one of several template DNAs, or 10 mM Tris-HCl (pH 8), 0.1 mM EDTA for non-template controls (NTC), in a total volume of 25 μl.

    Techniques: DNA Extraction

    Antibody titers to HIV-1 gp160 subtype (B) following pCMV-EnvABC and T20 peptide immunization. Six mice in each group were immunized with pCMV-EnvABC (20 µg) with T20 peptide (2 µg). Immunogens were given intramuscularly (im), intradermally (id) or intranasally (ina). Sera or tissue antibody levels were assayed by ELISA. Median IgG titers and ranges are shown.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Immunization with HIV-1 envelope T20-encoding DNA vaccines elicits cross-clade neutralizing antibody responses

    doi: 10.1080/21645515.2017.1338546

    Figure Lengend Snippet: Antibody titers to HIV-1 gp160 subtype (B) following pCMV-EnvABC and T20 peptide immunization. Six mice in each group were immunized with pCMV-EnvABC (20 µg) with T20 peptide (2 µg). Immunogens were given intramuscularly (im), intradermally (id) or intranasally (ina). Sera or tissue antibody levels were assayed by ELISA. Median IgG titers and ranges are shown.

    Article Snippet: Minicircle MC-T20 ( ): The nucleotide (nt) sequence for the T20 peptide with a secretion signal (ss) from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Immunomedics Inc., Morris Plains, NJ, USA) was cloned into the pCMV-T20 plasmid (see pCMV-EnvABC ) using Fse I and EcoRV restriction sites, by enzymes from New England Biolabs (R0588S and R0195S, NEB).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Schematic overview of T20 constructs. New constructs representing MC-T20, pCMV-T20, pCMVEE-T20 and the T20 peptide amino acid sequence with the 2F5 epitope in bold letters.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Immunization with HIV-1 envelope T20-encoding DNA vaccines elicits cross-clade neutralizing antibody responses

    doi: 10.1080/21645515.2017.1338546

    Figure Lengend Snippet: Schematic overview of T20 constructs. New constructs representing MC-T20, pCMV-T20, pCMVEE-T20 and the T20 peptide amino acid sequence with the 2F5 epitope in bold letters.

    Article Snippet: Minicircle MC-T20 ( ): The nucleotide (nt) sequence for the T20 peptide with a secretion signal (ss) from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Immunomedics Inc., Morris Plains, NJ, USA) was cloned into the pCMV-T20 plasmid (see pCMV-EnvABC ) using Fse I and EcoRV restriction sites, by enzymes from New England Biolabs (R0588S and R0195S, NEB).

    Techniques: Construct, Sequencing

    A-D. Neutralization titers to HIV-1 of subtypes A, B, (C) and CRF_AE. Five mice in each group were immunized with pCMV-EnvABC (20 µg) and MC-T20 20 µg; pCMV-EnvABC (20 µg) and pCMV-T20 (20 µg); pCMV-EnvABC (20 µg) and T20 peptide (10 µg); pCMV-EnvABC (20 µg) and non-coding pCMV-DNA (20 µg); or non-coding pCMV-DNA (40 µg). The p24 ELISA was used to detect subtype A, B and C viral antigen inhibition, the RT assay to detect CRF_AE viral antigen inhibition. (See also Table S1, Group III.)

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Immunization with HIV-1 envelope T20-encoding DNA vaccines elicits cross-clade neutralizing antibody responses

    doi: 10.1080/21645515.2017.1338546

    Figure Lengend Snippet: A-D. Neutralization titers to HIV-1 of subtypes A, B, (C) and CRF_AE. Five mice in each group were immunized with pCMV-EnvABC (20 µg) and MC-T20 20 µg; pCMV-EnvABC (20 µg) and pCMV-T20 (20 µg); pCMV-EnvABC (20 µg) and T20 peptide (10 µg); pCMV-EnvABC (20 µg) and non-coding pCMV-DNA (20 µg); or non-coding pCMV-DNA (40 µg). The p24 ELISA was used to detect subtype A, B and C viral antigen inhibition, the RT assay to detect CRF_AE viral antigen inhibition. (See also Table S1, Group III.)

    Article Snippet: Minicircle MC-T20 ( ): The nucleotide (nt) sequence for the T20 peptide with a secretion signal (ss) from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Immunomedics Inc., Morris Plains, NJ, USA) was cloned into the pCMV-T20 plasmid (see pCMV-EnvABC ) using Fse I and EcoRV restriction sites, by enzymes from New England Biolabs (R0588S and R0195S, NEB).

    Techniques: Neutralization, Mouse Assay, Enzyme-linked Immunosorbent Assay, Inhibition

    A-B. Anti-gp140 (C) and anti-T20 antibody titers following immunizations with pCMV-EnvABC with and without MC-T20 . Five mice in each group were immunized with pCMV-EnvABC (20 µg) with MC-T20 (20 µg); or pCMV-EnvABC (20 µg) with pCMV-DNA (20 µg). Sera were taken and antibodies against gp140 C and T20 peptide measured by ELISA. (See also Table S1, Group II.)

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Immunization with HIV-1 envelope T20-encoding DNA vaccines elicits cross-clade neutralizing antibody responses

    doi: 10.1080/21645515.2017.1338546

    Figure Lengend Snippet: A-B. Anti-gp140 (C) and anti-T20 antibody titers following immunizations with pCMV-EnvABC with and without MC-T20 . Five mice in each group were immunized with pCMV-EnvABC (20 µg) with MC-T20 (20 µg); or pCMV-EnvABC (20 µg) with pCMV-DNA (20 µg). Sera were taken and antibodies against gp140 C and T20 peptide measured by ELISA. (See also Table S1, Group II.)

    Article Snippet: Minicircle MC-T20 ( ): The nucleotide (nt) sequence for the T20 peptide with a secretion signal (ss) from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Immunomedics Inc., Morris Plains, NJ, USA) was cloned into the pCMV-T20 plasmid (see pCMV-EnvABC ) using Fse I and EcoRV restriction sites, by enzymes from New England Biolabs (R0588S and R0195S, NEB).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    A-D. Neutralization titers to HIV-1 of subtypes (B) and CRF_AE. Five mice in each group were immunized with pCMV-EnvABC (20 µg) with MC-T20 (20 µg); or MC-T20 (20 µg); or pCMVEE-T20 (2 µg). Sera were taken, divided into pools of sera with higher ELISA titers (A and B) and lower binding titers (C and D), and neutralization performed. The p24 ELISA was used to detect subtype B viral antigen inhibition, the RT assay to detect CRF_AE viral antigen inhibition. Positive sera consisted of hmAb 2F5 (inverted triangles) and 4E10 (diamonds). (See also Table S1, Group II.)

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Immunization with HIV-1 envelope T20-encoding DNA vaccines elicits cross-clade neutralizing antibody responses

    doi: 10.1080/21645515.2017.1338546

    Figure Lengend Snippet: A-D. Neutralization titers to HIV-1 of subtypes (B) and CRF_AE. Five mice in each group were immunized with pCMV-EnvABC (20 µg) with MC-T20 (20 µg); or MC-T20 (20 µg); or pCMVEE-T20 (2 µg). Sera were taken, divided into pools of sera with higher ELISA titers (A and B) and lower binding titers (C and D), and neutralization performed. The p24 ELISA was used to detect subtype B viral antigen inhibition, the RT assay to detect CRF_AE viral antigen inhibition. Positive sera consisted of hmAb 2F5 (inverted triangles) and 4E10 (diamonds). (See also Table S1, Group II.)

    Article Snippet: Minicircle MC-T20 ( ): The nucleotide (nt) sequence for the T20 peptide with a secretion signal (ss) from the V-J2-C region of the mouse Ig kappa-chain for efficient secretion of recombinant proteins (Immunomedics Inc., Morris Plains, NJ, USA) was cloned into the pCMV-T20 plasmid (see pCMV-EnvABC ) using Fse I and EcoRV restriction sites, by enzymes from New England Biolabs (R0588S and R0195S, NEB).

    Techniques: Neutralization, Mouse Assay, Enzyme-linked Immunosorbent Assay, Binding Assay, Inhibition

    Rate dependence of linear QPA on the length of AT-rich segment. (A) Targets and primer sequences; targets 1, 2 and 3 contain 34-nt, 20-nt and 5-nt AT-rich segments, respectively. (B) Typical dGTP-QPA (black) and dNTP-QPA profiles (colored) obtained using 0.5 μM primer, 10 nM target 2, 100 μM dGTP or 800 μM dNTP, 0.08 U/uL Bst 2.0 polymerase in 10 mM KCl, 40 mM CsCl, 2 mM MgCl 2 and 10 mM Tris-HCl, pH 8.7 at 62 °C. Dashed line corresponds to no-template control (NTC). (C) Temperature dependence of the rate of linear QPA using target 1 (blue), target 2 (red) and target 3 (black).

    Journal: Analytical methods : advancing methods and applications

    Article Title: Isothermal amplification of long DNA segments by quadruplex priming amplification

    doi: 10.1039/C8AY00843D

    Figure Lengend Snippet: Rate dependence of linear QPA on the length of AT-rich segment. (A) Targets and primer sequences; targets 1, 2 and 3 contain 34-nt, 20-nt and 5-nt AT-rich segments, respectively. (B) Typical dGTP-QPA (black) and dNTP-QPA profiles (colored) obtained using 0.5 μM primer, 10 nM target 2, 100 μM dGTP or 800 μM dNTP, 0.08 U/uL Bst 2.0 polymerase in 10 mM KCl, 40 mM CsCl, 2 mM MgCl 2 and 10 mM Tris-HCl, pH 8.7 at 62 °C. Dashed line corresponds to no-template control (NTC). (C) Temperature dependence of the rate of linear QPA using target 1 (blue), target 2 (red) and target 3 (black).

    Article Snippet: DNA polymerases ( Vent (exo-) and Bst 2.0), dNTPs and isothermal buffer (10 mM (NH4 )2 SO4 , 50 mM KCl, 2 mM MgSO4 , 0.1% Tween® 20, 20 mM Tris-HCl, pH 8.8) were purchased from New England BioLabs.

    Techniques: