bst large fragment polymerase  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Name:
    Bst DNA Polymerase Lg Frag
    Description:
    Bst DNA Polymerase Lg Frag 8 000 units
    Catalog Number:
    m0275l
    Price:
    277
    Size:
    8 000 units
    Category:
    Thermostable DNA Polymerases
    Buy from Supplier


    Structured Review

    New England Biolabs bst large fragment polymerase
    Bst DNA Polymerase Lg Frag
    Bst DNA Polymerase Lg Frag 8 000 units
    https://www.bioz.com/result/bst large fragment polymerase/product/New England Biolabs
    Average 90 stars, based on 106 article reviews
    Price from $9.99 to $1999.99
    bst large fragment polymerase - by Bioz Stars, 2020-01
    90/100 stars

    Images

    Related Articles

    Clone Assay:

    Article Title: Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR
    Article Snippet: .. The RT-LAMP reactions were carried out in a 12.5-μl reaction volume containing the primer concentrations indicated in ; a 1× concentration of ThermoPol buffer (New England BioLabs [NEB], Ipswich, MA), 8 mM magnesium sulfate (Invitrogen), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 1.4 mM each deoxynucleoside triphosphate (dNTP) (Qiagen, Hilden, Germany), 0.25 μl ResoLight dye (Roche Diagnostics), 3 U of Bst DNA polymerase (large fragment; NEB), and 3 U of cloned avian myeloblastosis virus (AMV) reverse transcriptase (Invitrogen). ..

    Amplification:

    Article Title: Loop-mediated isothermal amplification (RT-LAMP): a new approach for the detection of foot-and-mouth disease virus and its sero-types in Pakistan
    Article Snippet: RT-LAMP The RT-LAMP was carried out in a volume of 25 µL reaction mixture containing 2.5 µL ThermoPol reaction buffer (New England Bio Labs Inc., Beverly, MA, USA), 1 µL MgSO4 (100 mM), 2 µL dNTP set (Fermentas, 10 mM), 5 µL betaine (Sigma-Aldrich, St. Louis, MO, USA, 5 M), 2 µL nuclease-free water, 1 µL hydroxy naphthol blue (Sigma Aldrich, 3 mM), 1 µL Bst DNA polymerase Lg fragment (New England Bio Labs Inc., Beverly, MA, USA), 0.2 µL AMV reverse transcriptase (New England Bio Labs Inc., Beverly, MA, USA), 0.1 µL forward and backward outer primers, each with a 5 pmol concentration, 1 µL forward and backward inner primers each with a 50 pmol concentration, 0.5 µL forward and backward loop primers each with a 25 pmol concentration and 2 µL of extracted RNA from the samples (with a 4 µg/µL concentration). .. The FMDV specific primers used in the RT-LAMP for amplification of FMDV RNA are given in .

    Article Title: Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay
    Article Snippet: The optimum reagent concentrations in the LAMP reactions mixes were 1× Thermopol buffer (New England Biolabs, Hitchin, UK), 3 μM internal primers, 0.6 μM external primers, 1 mM MgSO4 (New England Biolabs, Hitchin, UK), 0.3 mM dNTPs (Sigma-Aldrich, Dorset, UK), 1 M betaine (Sigma-Aldrich, Dorset, UK), 16 U of Bst DNA polymerase (Large fragment: (New England Biolabs, Hitchin, UK), along with the addition of 2 μl of target DNA. .. Restriction enzyme digests were performed on the P32 LAMP amplification products to confirm that the correct region of the CaPV genome had been amplified.

    Article Title: Real-Time Reverse Transcription Loop-Mediated Isothermal Amplification for Rapid Detection of West Nile Virus
    Article Snippet: .. The RT-LAMP reaction was carried out in a 25-μl total reaction mixture volume with a Loopamp DNA amplification kit (Eiken Chemical Co. Ltd.) containing 50 pmol each of inner primers FIP and BIP, 5 pmol each of outer primers F3 and B3, 25 pmol each of loop primers loop F and loop B, 1,400 μM each deoxynucleoside triphosphate, 0.6 M betaine (Sigma Chemical Co., St. Louis, Mo.), 40 mM Tris-HCl (pH 8.8), 20 mM KCl, 20 mM (NH4 )2 SO4 , 8 mM MgSO4 , 0.1% Triton X-100, 0.125 U of avian myeloblastosis virus (AMV) RTase (Invitrogen), 8 U of Bst DNA polymerase (large fragment; New England Biolabs), and the specified amounts of target RNA. .. The mixture was incubated at 63°C for 60 min in a heating block and then heated at 80°C for 2 min to terminate the reaction.

    Article Title: A loop-mediated isothermal amplification assay for the visual detection of duck circovirus
    Article Snippet: Optimization of the DuCV LAMP conditions The DuCV LAMP assay was performed in tubes containing 10× Thermopol® Reaction Buffer (New England Biolabs, Beijing, China), Bst DNA polymerase (large fragment; New England Biolabs), dNTPs (Takara, Dalian, China), primers, betaine (Sigma–Aldrich), MgSO4 (Sigma–Aldrich), calcein (International Laboratory, USA), MnCl2 (International Laboratory, USA), template DNA/RNA and nuclease-free water. .. Based on the previous studies, different combinations of various concentrations of each component (dNTPs (0.4 mmol/L ~1.6 mmol/L), betaine (0.8 mmol/L ~1.4 mmol/L), MgSO4 (2 mmol/L ~9 mmol/L) were tested for amplification efficiency.

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: To the chamber, a 40 μl solution containing 0.8 units/μl of Bst polymerase (NEB, Cat. No. M0275S), 100 μM dNTP (NEB, Cat. No. N0447S), and the relevant primer mixes typically at 100 nM each, in 1× ThermoPol reaction buffer, was added and incubated for 1 h at room temperature. .. This minimizes any false-positive PCR amplification by the recording primers.

    Article Title: Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR
    Article Snippet: Paragraph title: Loop-mediated isothermal amplification. ... The RT-LAMP reactions were carried out in a 12.5-μl reaction volume containing the primer concentrations indicated in ; a 1× concentration of ThermoPol buffer (New England BioLabs [NEB], Ipswich, MA), 8 mM magnesium sulfate (Invitrogen), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 1.4 mM each deoxynucleoside triphosphate (dNTP) (Qiagen, Hilden, Germany), 0.25 μl ResoLight dye (Roche Diagnostics), 3 U of Bst DNA polymerase (large fragment; NEB), and 3 U of cloned avian myeloblastosis virus (AMV) reverse transcriptase (Invitrogen).

    Article Title: Loop-Mediated Isothermal Amplification for Laboratory Confirmation of Buruli Ulcer Disease—Towards a Point-of-Care Test
    Article Snippet: Conventional IS2404 LAMP (cLAMP) protocol Each cLAMP reaction mix consisted of 1 μl Bst DNA polymerase (large fragment, 8 U/μl; New England Biolabs [NEB], Frankfurt am Main, Germany), 1.0 μl dNTP mix (2 mM each, Merck), 1.0 μl of primers MU2-F3 and MU2-B3 (5 pmol/μl) and 2.0 μl of primers MU2-FIP and MU2-BIP (10 pmol/μl), respectively (TibMolBiol, Berlin, Germany), 2.0 μl betaine (5 M; Sigma-Aldrich, Taufkirchen, Germany), 2.5 μl 10-fold Thermopol buffer for Bst DNA polymerase (NEB) and 11.5 μl molecular grade H2 O (Carl Roth). .. Following the addition of 1 μl DNA extract (template), cLAMP reactions (final volume: 25 μl) were carried out in 1.5 ml SafeSeal reaction tubes (Sarstedt, Nümbrecht, Germany) at 65°C for 60 minutes in a conventional thermoblock (HLC Thermomixer MKR 13, HLC BioTech, Bovenden, Germany) and a final step at 80°C for 10 minutes terminated the amplification.

    Article Title: Thirty-minute screening of antibiotic resistance genes in bacterial isolates with minimal sample preparation in static self-dispensing 64 and 384 assay cards
    Article Snippet: Paragraph title: LAMP amplification ... Briefly, reaction mixtures contained 1× Bst DNA polymerase buffer (M0275L, New England Biolabs, Ipswich, MA, USA), 1.4 mM of each dNTP (10297018, Life Technologies, Grand Island, NY, USA), 800 mM betaine (B0300, Sigma Aldrich, St. Louis, MO, USA), 6 mM MgSO4 (B1003S, New England Biolabs, Ipswich, MA, USA), 1× primer mix (1.6 μM FIP and BIP, 800 nM LF and LB and 200 nM F3 and B3 primers), 1 mg/mL bovine serum albumin (B9000S, New England Biolabs, Ipswich, MA, USA), 20 μM SYTO-81 (S11362, Molecular Probes, Eugene, OR, USA [product not available anymore, can be replaced with SYTO-82, S11363, Life Technologies, Grand Island, NY, USA]), 0.2 % Pluronic F-68 (24040032, Life Technologies, Grand Island, NY, USA), and 0.64 U/μL Bst DNA polymerase (large fragment) (M0275L, New England Biolabs, Ipswich, MA, USA).

    Quantitative RT-PCR:

    Article Title: Loop-mediated isothermal amplification (RT-LAMP): a new approach for the detection of foot-and-mouth disease virus and its sero-types in Pakistan
    Article Snippet: The FMDV isolates were already confirmed by rRT-PCR (Reid et al., 2000 , 2002) and typed by indirect sandwich ELISA (Roeder and Smith, 1987 ; Ferris and Dawson, 1988 ). .. RT-LAMP The RT-LAMP was carried out in a volume of 25 µL reaction mixture containing 2.5 µL ThermoPol reaction buffer (New England Bio Labs Inc., Beverly, MA, USA), 1 µL MgSO4 (100 mM), 2 µL dNTP set (Fermentas, 10 mM), 5 µL betaine (Sigma-Aldrich, St. Louis, MO, USA, 5 M), 2 µL nuclease-free water, 1 µL hydroxy naphthol blue (Sigma Aldrich, 3 mM), 1 µL Bst DNA polymerase Lg fragment (New England Bio Labs Inc., Beverly, MA, USA), 0.2 µL AMV reverse transcriptase (New England Bio Labs Inc., Beverly, MA, USA), 0.1 µL forward and backward outer primers, each with a 5 pmol concentration, 1 µL forward and backward inner primers each with a 50 pmol concentration, 0.5 µL forward and backward loop primers each with a 25 pmol concentration and 2 µL of extracted RNA from the samples (with a 4 µg/µL concentration).

    Electrophoresis:

    Article Title: Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay
    Article Snippet: Optimisation of the Capripoxvirus LAMP assay In initial experiments, all three candidate LAMP assays that targeted the RNA polymerase subunit RPO30, DNA topoisomerase I and the P32 regions generated characteristic laddering patterns after agarose-gel electrophoresis (Figure A) and an increase in fluorescence in a real-time PCR machine (Figure B). .. The optimum reagent concentrations in the LAMP reactions mixes were 1× Thermopol buffer (New England Biolabs, Hitchin, UK), 3 μM internal primers, 0.6 μM external primers, 1 mM MgSO4 (New England Biolabs, Hitchin, UK), 0.3 mM dNTPs (Sigma-Aldrich, Dorset, UK), 1 M betaine (Sigma-Aldrich, Dorset, UK), 16 U of Bst DNA polymerase (Large fragment: (New England Biolabs, Hitchin, UK), along with the addition of 2 μl of target DNA.

    Sandwich ELISA:

    Article Title: Loop-mediated isothermal amplification (RT-LAMP): a new approach for the detection of foot-and-mouth disease virus and its sero-types in Pakistan
    Article Snippet: The FMDV isolates were already confirmed by rRT-PCR (Reid et al., 2000 , 2002) and typed by indirect sandwich ELISA (Roeder and Smith, 1987 ; Ferris and Dawson, 1988 ). .. RT-LAMP The RT-LAMP was carried out in a volume of 25 µL reaction mixture containing 2.5 µL ThermoPol reaction buffer (New England Bio Labs Inc., Beverly, MA, USA), 1 µL MgSO4 (100 mM), 2 µL dNTP set (Fermentas, 10 mM), 5 µL betaine (Sigma-Aldrich, St. Louis, MO, USA, 5 M), 2 µL nuclease-free water, 1 µL hydroxy naphthol blue (Sigma Aldrich, 3 mM), 1 µL Bst DNA polymerase Lg fragment (New England Bio Labs Inc., Beverly, MA, USA), 0.2 µL AMV reverse transcriptase (New England Bio Labs Inc., Beverly, MA, USA), 0.1 µL forward and backward outer primers, each with a 5 pmol concentration, 1 µL forward and backward inner primers each with a 50 pmol concentration, 0.5 µL forward and backward loop primers each with a 25 pmol concentration and 2 µL of extracted RNA from the samples (with a 4 µg/µL concentration).

    Incubation:

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: A volume of 9 μl of 1 mM DBCO-linker strands and 1 μl of 10× phosphate-buffered saline (pH 7.4) buffer were added to 20 μl of water containing 180 pmol azide-modified probes and incubated at room temperature for 12 h. The linked probes were purified by 8% denaturing polyacrylamide gel and quantified by NanoDrop UV–Vis Spectrophotometer (Thermo Scientific). .. This domain was subsequently extended with Bst large fragment polymerase (NEB M0275), followed by USER-mediated cleavage (Uracil-Specific Excision Reagent, NEB, M5505) of the 5′ uracil-containing domains and polyacrylamide gel purification.

    Article Title: Real-Time Reverse Transcription Loop-Mediated Isothermal Amplification for Rapid Detection of West Nile Virus
    Article Snippet: The RT-LAMP reaction was carried out in a 25-μl total reaction mixture volume with a Loopamp DNA amplification kit (Eiken Chemical Co. Ltd.) containing 50 pmol each of inner primers FIP and BIP, 5 pmol each of outer primers F3 and B3, 25 pmol each of loop primers loop F and loop B, 1,400 μM each deoxynucleoside triphosphate, 0.6 M betaine (Sigma Chemical Co., St. Louis, Mo.), 40 mM Tris-HCl (pH 8.8), 20 mM KCl, 20 mM (NH4 )2 SO4 , 8 mM MgSO4 , 0.1% Triton X-100, 0.125 U of avian myeloblastosis virus (AMV) RTase (Invitrogen), 8 U of Bst DNA polymerase (large fragment; New England Biolabs), and the specified amounts of target RNA. .. The mixture was incubated at 63°C for 60 min in a heating block and then heated at 80°C for 2 min to terminate the reaction.

    Article Title: A loop-mediated isothermal amplification assay for the visual detection of duck circovirus
    Article Snippet: Optimization of the DuCV LAMP conditions The DuCV LAMP assay was performed in tubes containing 10× Thermopol® Reaction Buffer (New England Biolabs, Beijing, China), Bst DNA polymerase (large fragment; New England Biolabs), dNTPs (Takara, Dalian, China), primers, betaine (Sigma–Aldrich), MgSO4 (Sigma–Aldrich), calcein (International Laboratory, USA), MnCl2 (International Laboratory, USA), template DNA/RNA and nuclease-free water. .. The amplification reaction was performed in a thermal block between 59°C to 65°C within 40 to 80 min, to ascertain the optimal incubation temperature and time.

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: .. To the chamber, a 40 μl solution containing 0.8 units/μl of Bst polymerase (NEB, Cat. No. M0275S), 100 μM dNTP (NEB, Cat. No. N0447S), and the relevant primer mixes typically at 100 nM each, in 1× ThermoPol reaction buffer, was added and incubated for 1 h at room temperature. .. After the recording reaction, the supernatant solution containing produced records was collected and the polymerase was heat inactivated by incubating the solution at 80 °C for 20 min. For samples used in the geometry studies (three-point and hexagonal grid patterns) and the state change study, before quenching the reaction, extra recording primers contained in the product solution were removed by mixing the solution with Exonuclease I (NEB, Cat. No. M0293S) at 9:1 volume ratio and incubating for 20 min at 37 °C.

    Article Title: Long‐fragment targeted capture for long‐read sequencing of plastomes
    Article Snippet: The mix is incubated for 30 min at room temperature (20°C), and the reaction is stopped with an AMPure XP 0.5× clean‐up step. .. Step 5: A nick fill‐in step is performed using 2 μL of 8 U/μL Bst DNA polymerase (#M0275; New England BioLabs), 3 μL of 10× ThermoPol Reaction Buffer (New England BioLabs), and 1.5 μL of 5 mM dNTPs in a final volume of 30 μL.

    Gel Purification:

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: .. This domain was subsequently extended with Bst large fragment polymerase (NEB M0275), followed by USER-mediated cleavage (Uracil-Specific Excision Reagent, NEB, M5505) of the 5′ uracil-containing domains and polyacrylamide gel purification. .. NUPACK simulations Expected reaction products at thermodynamic equilibrium were predicted using NUPACK web-based software ( nupack.org ), analysis option.

    High Performance Liquid Chromatography:

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: Both DNA strands were HPLC purified by the manufacturers. .. This domain was subsequently extended with Bst large fragment polymerase (NEB M0275), followed by USER-mediated cleavage (Uracil-Specific Excision Reagent, NEB, M5505) of the 5′ uracil-containing domains and polyacrylamide gel purification.

    Article Title: Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR
    Article Snippet: For both assays, FIP and BIP primers were purified by high-performance liquid chromatography (HPLC). .. The RT-LAMP reactions were carried out in a 12.5-μl reaction volume containing the primer concentrations indicated in ; a 1× concentration of ThermoPol buffer (New England BioLabs [NEB], Ipswich, MA), 8 mM magnesium sulfate (Invitrogen), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 1.4 mM each deoxynucleoside triphosphate (dNTP) (Qiagen, Hilden, Germany), 0.25 μl ResoLight dye (Roche Diagnostics), 3 U of Bst DNA polymerase (large fragment; NEB), and 3 U of cloned avian myeloblastosis virus (AMV) reverse transcriptase (Invitrogen).

    Flow Cytometry:

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: For recording reactions on origami, flow chambers were created by attaching a freshly cleaved mica (Ted Pella) piece to a channel slide system (ibidi, Sticky-Slide VI0.4, Cat. No. 80608), yielding a 30 μl chamber volume. .. To the chamber, a 40 μl solution containing 0.8 units/μl of Bst polymerase (NEB, Cat. No. M0275S), 100 μM dNTP (NEB, Cat. No. N0447S), and the relevant primer mixes typically at 100 nM each, in 1× ThermoPol reaction buffer, was added and incubated for 1 h at room temperature.

    Generated:

    Article Title: Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay
    Article Snippet: Optimisation of the Capripoxvirus LAMP assay In initial experiments, all three candidate LAMP assays that targeted the RNA polymerase subunit RPO30, DNA topoisomerase I and the P32 regions generated characteristic laddering patterns after agarose-gel electrophoresis (Figure A) and an increase in fluorescence in a real-time PCR machine (Figure B). .. The optimum reagent concentrations in the LAMP reactions mixes were 1× Thermopol buffer (New England Biolabs, Hitchin, UK), 3 μM internal primers, 0.6 μM external primers, 1 mM MgSO4 (New England Biolabs, Hitchin, UK), 0.3 mM dNTPs (Sigma-Aldrich, Dorset, UK), 1 M betaine (Sigma-Aldrich, Dorset, UK), 16 U of Bst DNA polymerase (Large fragment: (New England Biolabs, Hitchin, UK), along with the addition of 2 μl of target DNA.

    other:

    Article Title: SELMA: Selection with Modified Aptamers
    Article Snippet: ssDNA library Form C (see Basic Protocol 1) BST DNA polymerase, large fragment, with 10× ThermoPol reaction buffer (New England Biolabs, M0275) 10 mM EdUTP mix (see recipe) Liquid nitrogen Nitrogen gas Argon gas (optional) 100 mM HEPES-KOH, pH 8.0, with 0.5% (v/v) Triton X-100 50 mM azide sugar (see recipe) 10 mM CuSO4 (see recipe) 10 mM THPTA (see recipe) (+)-Sodium l -ascorbate Thermal cycler Two 50-ml, two-neck, pear-shaped flasks with ground-glass joints and pointed bottom (e.g., Chemglass, cat. no. CG-1558-14) Gas/vacuum manifold 500-μl tubes SpeedVac concentrator Rubber septum Additional reagents and equipment for desalting (see Basic Protocol 1)

    Sequencing:

    Article Title: Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR
    Article Snippet: Placement of a Loop-B primer was not possible due to low sequence conservation in the respective genomic region of the 5′ UTR. .. The RT-LAMP reactions were carried out in a 12.5-μl reaction volume containing the primer concentrations indicated in ; a 1× concentration of ThermoPol buffer (New England BioLabs [NEB], Ipswich, MA), 8 mM magnesium sulfate (Invitrogen), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 1.4 mM each deoxynucleoside triphosphate (dNTP) (Qiagen, Hilden, Germany), 0.25 μl ResoLight dye (Roche Diagnostics), 3 U of Bst DNA polymerase (large fragment; NEB), and 3 U of cloned avian myeloblastosis virus (AMV) reverse transcriptase (Invitrogen).

    Binding Assay:

    Article Title: Fabrication of DNA Polymer Brush Arrays by Destructive Micropatterning and Rolling-Circle Amplification
    Article Snippet: Bst DNA polymerase large fragment (Cat. # M0275L, New England Biolabs) was introduced at a concentration of 1 U · μL−1 in the polymerase buffer. .. The chip was cooled to 4 °C and a reaction mix containing 1 U · μL−1 Bst DNA polymerase, 100 μg · mL−1 BSA (Cat. # B9001S, New England Biolabs), 10−3 m each of dATP, dCTP, dGTP, and dTTP (Cat. #s 27-2050-02, 27-2060-02, 27-2070-02, and 27-2080-02, Amersham Biosciences), 4 × 10−6 m Escherichia coli single-stranded DNA binding protein (SSB, Cat. # 70032Z, United States Biochemicals) in a Bst buffer [0.020 m Tris-HCl, 10−3 m (NH4 )2 SO4 , 0.050 m KCl, 4 × 10−3 m MgSO4 , 0.1% Triton X-100, pH = 8.8 at 25 °C] was loaded.

    Fluorescence:

    Article Title: Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay
    Article Snippet: Optimisation of the Capripoxvirus LAMP assay In initial experiments, all three candidate LAMP assays that targeted the RNA polymerase subunit RPO30, DNA topoisomerase I and the P32 regions generated characteristic laddering patterns after agarose-gel electrophoresis (Figure A) and an increase in fluorescence in a real-time PCR machine (Figure B). .. The optimum reagent concentrations in the LAMP reactions mixes were 1× Thermopol buffer (New England Biolabs, Hitchin, UK), 3 μM internal primers, 0.6 μM external primers, 1 mM MgSO4 (New England Biolabs, Hitchin, UK), 0.3 mM dNTPs (Sigma-Aldrich, Dorset, UK), 1 M betaine (Sigma-Aldrich, Dorset, UK), 16 U of Bst DNA polymerase (Large fragment: (New England Biolabs, Hitchin, UK), along with the addition of 2 μl of target DNA.

    Isolation:

    Article Title: Efficient and Specific Detection of Salmonella in Food Samples Using a stn-Based Loop-Mediated Isothermal Amplification Method
    Article Snippet: LAMP Reaction and LAMP Product Detection The LAMP reaction was performed in a total volume of 25 μ L containing the following components (final concentration): 1.6 μ M each of FIP and BIP primers, 0.2 μ M each of F3 and B3 primers, 0.8 μ M each of LF and LB primers (in the same LAMP reaction), 1.6 mM of deoxyribonucleotide triphosphate mixture (dNTPs), 1 M betaine (Sigma, B2629, St. Louis, USA), 6 mM MgSO4 , 1x thermopol buffer (New England Biolabs, B9004S, Beverly, USA), 1 μ L (8 U) of B st DNA polymerase large fragment (New England Biolabs, M0275S, Beverly, USA), and 5 μ L of DNA template solution. .. Typhimurium ATCC 23566 cells or its isolated DNA (1 ng/reaction) was used as positive controls.

    Purification:

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: A volume of 9 μl of 1 mM DBCO-linker strands and 1 μl of 10× phosphate-buffered saline (pH 7.4) buffer were added to 20 μl of water containing 180 pmol azide-modified probes and incubated at room temperature for 12 h. The linked probes were purified by 8% denaturing polyacrylamide gel and quantified by NanoDrop UV–Vis Spectrophotometer (Thermo Scientific). .. This domain was subsequently extended with Bst large fragment polymerase (NEB M0275), followed by USER-mediated cleavage (Uracil-Specific Excision Reagent, NEB, M5505) of the 5′ uracil-containing domains and polyacrylamide gel purification.

    Article Title: Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR
    Article Snippet: For both assays, FIP and BIP primers were purified by high-performance liquid chromatography (HPLC). .. The RT-LAMP reactions were carried out in a 12.5-μl reaction volume containing the primer concentrations indicated in ; a 1× concentration of ThermoPol buffer (New England BioLabs [NEB], Ipswich, MA), 8 mM magnesium sulfate (Invitrogen), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 1.4 mM each deoxynucleoside triphosphate (dNTP) (Qiagen, Hilden, Germany), 0.25 μl ResoLight dye (Roche Diagnostics), 3 U of Bst DNA polymerase (large fragment; NEB), and 3 U of cloned avian myeloblastosis virus (AMV) reverse transcriptase (Invitrogen).

    Article Title: Long‐fragment targeted capture for long‐read sequencing of plastomes
    Article Snippet: A clean‐up step is performed with an AMPure XP 1× purification. .. Step 5: A nick fill‐in step is performed using 2 μL of 8 U/μL Bst DNA polymerase (#M0275; New England BioLabs), 3 μL of 10× ThermoPol Reaction Buffer (New England BioLabs), and 1.5 μL of 5 mM dNTPs in a final volume of 30 μL.

    Polymerase Chain Reaction:

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: To the chamber, a 40 μl solution containing 0.8 units/μl of Bst polymerase (NEB, Cat. No. M0275S), 100 μM dNTP (NEB, Cat. No. N0447S), and the relevant primer mixes typically at 100 nM each, in 1× ThermoPol reaction buffer, was added and incubated for 1 h at room temperature. .. This minimizes any false-positive PCR amplification by the recording primers.

    Blocking Assay:

    Article Title: Real-Time Reverse Transcription Loop-Mediated Isothermal Amplification for Rapid Detection of West Nile Virus
    Article Snippet: The RT-LAMP reaction was carried out in a 25-μl total reaction mixture volume with a Loopamp DNA amplification kit (Eiken Chemical Co. Ltd.) containing 50 pmol each of inner primers FIP and BIP, 5 pmol each of outer primers F3 and B3, 25 pmol each of loop primers loop F and loop B, 1,400 μM each deoxynucleoside triphosphate, 0.6 M betaine (Sigma Chemical Co., St. Louis, Mo.), 40 mM Tris-HCl (pH 8.8), 20 mM KCl, 20 mM (NH4 )2 SO4 , 8 mM MgSO4 , 0.1% Triton X-100, 0.125 U of avian myeloblastosis virus (AMV) RTase (Invitrogen), 8 U of Bst DNA polymerase (large fragment; New England Biolabs), and the specified amounts of target RNA. .. The mixture was incubated at 63°C for 60 min in a heating block and then heated at 80°C for 2 min to terminate the reaction.

    Article Title: A loop-mediated isothermal amplification assay for the visual detection of duck circovirus
    Article Snippet: Optimization of the DuCV LAMP conditions The DuCV LAMP assay was performed in tubes containing 10× Thermopol® Reaction Buffer (New England Biolabs, Beijing, China), Bst DNA polymerase (large fragment; New England Biolabs), dNTPs (Takara, Dalian, China), primers, betaine (Sigma–Aldrich), MgSO4 (Sigma–Aldrich), calcein (International Laboratory, USA), MnCl2 (International Laboratory, USA), template DNA/RNA and nuclease-free water. .. The amplification reaction was performed in a thermal block between 59°C to 65°C within 40 to 80 min, to ascertain the optimal incubation temperature and time.

    Chromatin Immunoprecipitation:

    Article Title: Fabrication of DNA Polymer Brush Arrays by Destructive Micropatterning and Rolling-Circle Amplification
    Article Snippet: A solution containing 20 n m of Circle1 or Circle2 in a 2X SSC solution was then introduced and the chip was heated to 75 °C for 2 min and cooled to 55 °C at 2 °C · min−1 . .. Bst DNA polymerase large fragment (Cat. # M0275L, New England Biolabs) was introduced at a concentration of 1 U · μL−1 in the polymerase buffer.

    Software:

    Article Title: Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR
    Article Snippet: The final primer set ( and ) was constructed by using the LAMP primer design software Primer Explorer V4 ( ). .. The RT-LAMP reactions were carried out in a 12.5-μl reaction volume containing the primer concentrations indicated in ; a 1× concentration of ThermoPol buffer (New England BioLabs [NEB], Ipswich, MA), 8 mM magnesium sulfate (Invitrogen), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 1.4 mM each deoxynucleoside triphosphate (dNTP) (Qiagen, Hilden, Germany), 0.25 μl ResoLight dye (Roche Diagnostics), 3 U of Bst DNA polymerase (large fragment; NEB), and 3 U of cloned avian myeloblastosis virus (AMV) reverse transcriptase (Invitrogen).

    Real-time Polymerase Chain Reaction:

    Article Title: Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay
    Article Snippet: Optimisation of the Capripoxvirus LAMP assay In initial experiments, all three candidate LAMP assays that targeted the RNA polymerase subunit RPO30, DNA topoisomerase I and the P32 regions generated characteristic laddering patterns after agarose-gel electrophoresis (Figure A) and an increase in fluorescence in a real-time PCR machine (Figure B). .. The optimum reagent concentrations in the LAMP reactions mixes were 1× Thermopol buffer (New England Biolabs, Hitchin, UK), 3 μM internal primers, 0.6 μM external primers, 1 mM MgSO4 (New England Biolabs, Hitchin, UK), 0.3 mM dNTPs (Sigma-Aldrich, Dorset, UK), 1 M betaine (Sigma-Aldrich, Dorset, UK), 16 U of Bst DNA polymerase (Large fragment: (New England Biolabs, Hitchin, UK), along with the addition of 2 μl of target DNA.

    Article Title: Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR
    Article Snippet: The RT-LAMP reactions were carried out in a 12.5-μl reaction volume containing the primer concentrations indicated in ; a 1× concentration of ThermoPol buffer (New England BioLabs [NEB], Ipswich, MA), 8 mM magnesium sulfate (Invitrogen), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 1.4 mM each deoxynucleoside triphosphate (dNTP) (Qiagen, Hilden, Germany), 0.25 μl ResoLight dye (Roche Diagnostics), 3 U of Bst DNA polymerase (large fragment; NEB), and 3 U of cloned avian myeloblastosis virus (AMV) reverse transcriptase (Invitrogen). .. For the final assays, amplification was performed on the Eco real-time PCR system (amplifa Labortechnik GmBH), using 60 cycles of 1 min at 63°C followed by a standard melting curve analysis.

    RNA Extraction:

    Article Title: Loop-mediated isothermal amplification (RT-LAMP): a new approach for the detection of foot-and-mouth disease virus and its sero-types in Pakistan
    Article Snippet: RNA extraction FMDV specific RNA was extracted from the above mentioned viruses using QIAamp® Viral RNA Minikit (Qiagen, GmBh, Germany) following the manufacturer’s instructions. .. RT-LAMP The RT-LAMP was carried out in a volume of 25 µL reaction mixture containing 2.5 µL ThermoPol reaction buffer (New England Bio Labs Inc., Beverly, MA, USA), 1 µL MgSO4 (100 mM), 2 µL dNTP set (Fermentas, 10 mM), 5 µL betaine (Sigma-Aldrich, St. Louis, MO, USA, 5 M), 2 µL nuclease-free water, 1 µL hydroxy naphthol blue (Sigma Aldrich, 3 mM), 1 µL Bst DNA polymerase Lg fragment (New England Bio Labs Inc., Beverly, MA, USA), 0.2 µL AMV reverse transcriptase (New England Bio Labs Inc., Beverly, MA, USA), 0.1 µL forward and backward outer primers, each with a 5 pmol concentration, 1 µL forward and backward inner primers each with a 50 pmol concentration, 0.5 µL forward and backward loop primers each with a 25 pmol concentration and 2 µL of extracted RNA from the samples (with a 4 µg/µL concentration).

    Agarose Gel Electrophoresis:

    Article Title: Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay
    Article Snippet: Optimisation of the Capripoxvirus LAMP assay In initial experiments, all three candidate LAMP assays that targeted the RNA polymerase subunit RPO30, DNA topoisomerase I and the P32 regions generated characteristic laddering patterns after agarose-gel electrophoresis (Figure A) and an increase in fluorescence in a real-time PCR machine (Figure B). .. The optimum reagent concentrations in the LAMP reactions mixes were 1× Thermopol buffer (New England Biolabs, Hitchin, UK), 3 μM internal primers, 0.6 μM external primers, 1 mM MgSO4 (New England Biolabs, Hitchin, UK), 0.3 mM dNTPs (Sigma-Aldrich, Dorset, UK), 1 M betaine (Sigma-Aldrich, Dorset, UK), 16 U of Bst DNA polymerase (Large fragment: (New England Biolabs, Hitchin, UK), along with the addition of 2 μl of target DNA.

    Spectrophotometry:

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: A volume of 9 μl of 1 mM DBCO-linker strands and 1 μl of 10× phosphate-buffered saline (pH 7.4) buffer were added to 20 μl of water containing 180 pmol azide-modified probes and incubated at room temperature for 12 h. The linked probes were purified by 8% denaturing polyacrylamide gel and quantified by NanoDrop UV–Vis Spectrophotometer (Thermo Scientific). .. This domain was subsequently extended with Bst large fragment polymerase (NEB M0275), followed by USER-mediated cleavage (Uracil-Specific Excision Reagent, NEB, M5505) of the 5′ uracil-containing domains and polyacrylamide gel purification.

    Produced:

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: To the chamber, a 40 μl solution containing 0.8 units/μl of Bst polymerase (NEB, Cat. No. M0275S), 100 μM dNTP (NEB, Cat. No. N0447S), and the relevant primer mixes typically at 100 nM each, in 1× ThermoPol reaction buffer, was added and incubated for 1 h at room temperature. .. After the recording reaction, the supernatant solution containing produced records was collected and the polymerase was heat inactivated by incubating the solution at 80 °C for 20 min. For samples used in the geometry studies (three-point and hexagonal grid patterns) and the state change study, before quenching the reaction, extra recording primers contained in the product solution were removed by mixing the solution with Exonuclease I (NEB, Cat. No. M0293S) at 9:1 volume ratio and incubating for 20 min at 37 °C.

    Concentration Assay:

    Article Title: Loop-mediated isothermal amplification (RT-LAMP): a new approach for the detection of foot-and-mouth disease virus and its sero-types in Pakistan
    Article Snippet: .. RT-LAMP The RT-LAMP was carried out in a volume of 25 µL reaction mixture containing 2.5 µL ThermoPol reaction buffer (New England Bio Labs Inc., Beverly, MA, USA), 1 µL MgSO4 (100 mM), 2 µL dNTP set (Fermentas, 10 mM), 5 µL betaine (Sigma-Aldrich, St. Louis, MO, USA, 5 M), 2 µL nuclease-free water, 1 µL hydroxy naphthol blue (Sigma Aldrich, 3 mM), 1 µL Bst DNA polymerase Lg fragment (New England Bio Labs Inc., Beverly, MA, USA), 0.2 µL AMV reverse transcriptase (New England Bio Labs Inc., Beverly, MA, USA), 0.1 µL forward and backward outer primers, each with a 5 pmol concentration, 1 µL forward and backward inner primers each with a 50 pmol concentration, 0.5 µL forward and backward loop primers each with a 25 pmol concentration and 2 µL of extracted RNA from the samples (with a 4 µg/µL concentration). .. The FMDV specific primers used in the RT-LAMP for amplification of FMDV RNA are given in .

    Article Title: Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay
    Article Snippet: The optimum reagent concentrations in the LAMP reactions mixes were 1× Thermopol buffer (New England Biolabs, Hitchin, UK), 3 μM internal primers, 0.6 μM external primers, 1 mM MgSO4 (New England Biolabs, Hitchin, UK), 0.3 mM dNTPs (Sigma-Aldrich, Dorset, UK), 1 M betaine (Sigma-Aldrich, Dorset, UK), 16 U of Bst DNA polymerase (Large fragment: (New England Biolabs, Hitchin, UK), along with the addition of 2 μl of target DNA. .. Using the P32 primer set, loop primers were added to the reaction at an optimal concentration of 4 μM.

    Article Title: Efficient and Specific Detection of Salmonella in Food Samples Using a stn-Based Loop-Mediated Isothermal Amplification Method
    Article Snippet: .. LAMP Reaction and LAMP Product Detection The LAMP reaction was performed in a total volume of 25 μ L containing the following components (final concentration): 1.6 μ M each of FIP and BIP primers, 0.2 μ M each of F3 and B3 primers, 0.8 μ M each of LF and LB primers (in the same LAMP reaction), 1.6 mM of deoxyribonucleotide triphosphate mixture (dNTPs), 1 M betaine (Sigma, B2629, St. Louis, USA), 6 mM MgSO4 , 1x thermopol buffer (New England Biolabs, B9004S, Beverly, USA), 1 μ L (8 U) of B st DNA polymerase large fragment (New England Biolabs, M0275S, Beverly, USA), and 5 μ L of DNA template solution. ..

    Article Title: Fabrication of DNA Polymer Brush Arrays by Destructive Micropatterning and Rolling-Circle Amplification
    Article Snippet: .. Bst DNA polymerase large fragment (Cat. # M0275L, New England Biolabs) was introduced at a concentration of 1 U · μL−1 in the polymerase buffer. .. The chip was then heated to 50 °C for 15 min to allow the polymerases to bind to the primed circular DNA molecules.

    Article Title: A DNA nanoscope via auto-cycling proximity recording
    Article Snippet: After washing the chamber three times with 60 μl of TAE/Mg buffer by adding the buffer to one reservoir (inlet) and subsequently taking out the same volume from the other side (outlet), a 30 μl solution containing origami at 50 pM concentration was introduced to the chamber. (In the first washing round, ~30 μl of buffer occupies the chamber and only ~30 μl of extra buffer comes out.) .. To the chamber, a 40 μl solution containing 0.8 units/μl of Bst polymerase (NEB, Cat. No. M0275S), 100 μM dNTP (NEB, Cat. No. N0447S), and the relevant primer mixes typically at 100 nM each, in 1× ThermoPol reaction buffer, was added and incubated for 1 h at room temperature.

    Article Title: Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR
    Article Snippet: .. The RT-LAMP reactions were carried out in a 12.5-μl reaction volume containing the primer concentrations indicated in ; a 1× concentration of ThermoPol buffer (New England BioLabs [NEB], Ipswich, MA), 8 mM magnesium sulfate (Invitrogen), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 1.4 mM each deoxynucleoside triphosphate (dNTP) (Qiagen, Hilden, Germany), 0.25 μl ResoLight dye (Roche Diagnostics), 3 U of Bst DNA polymerase (large fragment; NEB), and 3 U of cloned avian myeloblastosis virus (AMV) reverse transcriptase (Invitrogen). ..

    Article Title: Long‐fragment targeted capture for long‐read sequencing of plastomes
    Article Snippet: DNA concentration is checked on NanoQuant Infinite M200 to ensure enough material is available for downstream steps. .. Step 5: A nick fill‐in step is performed using 2 μL of 8 U/μL Bst DNA polymerase (#M0275; New England BioLabs), 3 μL of 10× ThermoPol Reaction Buffer (New England BioLabs), and 1.5 μL of 5 mM dNTPs in a final volume of 30 μL.

    Construct:

    Article Title: Rapid Genome Detection of Schmallenberg Virus and Bovine Viral Diarrhea Virus by Use of Isothermal Amplification Methods and High-Speed Real-Time Reverse Transcriptase PCR
    Article Snippet: The final primer set ( and ) was constructed by using the LAMP primer design software Primer Explorer V4 ( ). .. The RT-LAMP reactions were carried out in a 12.5-μl reaction volume containing the primer concentrations indicated in ; a 1× concentration of ThermoPol buffer (New England BioLabs [NEB], Ipswich, MA), 8 mM magnesium sulfate (Invitrogen), 0.8 M betaine (Sigma-Aldrich, St. Louis, MO), 1.4 mM each deoxynucleoside triphosphate (dNTP) (Qiagen, Hilden, Germany), 0.25 μl ResoLight dye (Roche Diagnostics), 3 U of Bst DNA polymerase (large fragment; NEB), and 3 U of cloned avian myeloblastosis virus (AMV) reverse transcriptase (Invitrogen).

    Lamp Assay:

    Article Title: Loop-mediated isothermal amplification (LAMP) assays for the species-specific detection of Eimeria that infect chickens
    Article Snippet: .. LAMP assay Each LAMP reaction was performed in a final volume of 25 μl containing 8 U Bst DNA polymerase (large fragment; New England Biolabs) in 1 × ThermoPol Reaction buffer (20 mM Tris-HCl, 10 mM KCl, 10 mM (NH4 )2 SO4 , 2 mM MgSO4 and 0.1% Triton X-100; New England Biolabs) supplemented with 2 mM MgCl2 , 1 M betaine and 400 μM of each dNTP. ..

    Article Title: Detection of capripoxvirus DNA using a novel loop-mediated isothermal amplification assay
    Article Snippet: Paragraph title: Optimisation of the Capripoxvirus LAMP assay ... The optimum reagent concentrations in the LAMP reactions mixes were 1× Thermopol buffer (New England Biolabs, Hitchin, UK), 3 μM internal primers, 0.6 μM external primers, 1 mM MgSO4 (New England Biolabs, Hitchin, UK), 0.3 mM dNTPs (Sigma-Aldrich, Dorset, UK), 1 M betaine (Sigma-Aldrich, Dorset, UK), 16 U of Bst DNA polymerase (Large fragment: (New England Biolabs, Hitchin, UK), along with the addition of 2 μl of target DNA.

    Article Title: A loop-mediated isothermal amplification assay for the visual detection of duck circovirus
    Article Snippet: .. Optimization of the DuCV LAMP conditions The DuCV LAMP assay was performed in tubes containing 10× Thermopol® Reaction Buffer (New England Biolabs, Beijing, China), Bst DNA polymerase (large fragment; New England Biolabs), dNTPs (Takara, Dalian, China), primers, betaine (Sigma–Aldrich), MgSO4 (Sigma–Aldrich), calcein (International Laboratory, USA), MnCl2 (International Laboratory, USA), template DNA/RNA and nuclease-free water. .. Based on the previous studies, different combinations of various concentrations of each component (dNTPs (0.4 mmol/L ~1.6 mmol/L), betaine (0.8 mmol/L ~1.4 mmol/L), MgSO4 (2 mmol/L ~9 mmol/L) were tested for amplification efficiency.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    New England Biolabs bst dna polymerase lg frag
    Bst Dna Polymerase Lg Frag, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bst dna polymerase lg frag/product/New England Biolabs
    Average 90 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    bst dna polymerase lg frag - by Bioz Stars, 2020-01
    90/100 stars
      Buy from Supplier

    Image Search Results