reaction buffer  (New England Biolabs)


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  • 99
    Name:
    Q5 Reaction Buffer Pack
    Description:
    Q5 Reaction Buffer Pack 6 0 ml
    Catalog Number:
    b9027s
    Price:
    28
    Size:
    6 0 ml
    Category:
    Buffers
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    Structured Review

    New England Biolabs reaction buffer
    Q5 Reaction Buffer Pack
    Q5 Reaction Buffer Pack 6 0 ml
    https://www.bioz.com/result/reaction buffer/product/New England Biolabs
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    reaction buffer - by Bioz Stars, 2020-09
    99/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: Genetic characterization of fall armyworm infesting South Africa and India indicate recent introduction from a common source population
    Article Snippet: .. PCR amplification for all segments was performed in a 30-μl reaction mix containing 3 μl 10X manufacturer’s reaction buffer, 1 μl 10mM dNTP, 0.5 μl 20-μM primer mix, 1 μl DNA template (between 0.05–0.5 μg), 0.5 unit Taq DNA polymerase (New England Biolabs, Beverly, MA). .. Typically 96 PCR amplifications were performed at the same time using either 0.2-ml tube strips or 96 well microtiter plates.

    Article Title: Comparative molecular analyses of invasive fall armyworm in Togo reveal strong similarities to populations from the eastern United States and the Greater Antilles
    Article Snippet: .. PCR amplification for all segments was performed in a 30-μl reaction mix containing 3 μl 10X manufacturer’s reaction buffer, 1 μl 10mM dNTP, 0.5 μl 20-μM primer mix, 1 μl DNA template (between 0.05–0.5 μg), 0.5 unit Taq DNA polymerase (New England Biolabs, Beverly, MA). .. Typically 96 PCR amplifications were performed at the same time using either 0.2-ml tube strips or 96 well microtiter plates.

    Incubation:

    Article Title: Identification and Characterization of Neuronal Mitogen-activated Protein Kinase Substrates Using a Specific Phosphomotif Antibody *Identification and Characterization of Neuronal Mitogen-activated Protein Kinase Substrates Using a Specific Phosphomotif Antibody * S⃞
    Article Snippet: .. Purified protein on beads was incubated at 30 °C for 30 min in 20 μl of reaction buffer with 5 μCi of [γ-32 P]ATP, 100 μm ATP, and active purified ERK2 (100 units; New England Biolabs), JNK1 (200 ng; Millipore), or no kinase. .. Similarly PSD1 fraction (30 μg) was phosphorylated with active ERK2, JNK1, GSK-3β, CaMKII (100 units with Ca2+ /calmodulin; New England Biolabs), or cyclin-dependent kinase 2 (CDK2) (100 units with cyclin A; New England Biolabs) in the presence of 100 μm unlabeled ATP and detected by 5557 immunoblotting.

    Article Title: Characterization of the major formamidopyrimidine-DNA glycosylase homolog in Mycobacterium tuberculosis and its linkage to variable tandem repeats
    Article Snippet: .. The enzyme activities were assayed in a reaction buffer containing 70 mM 3-(N -morpholino) propane sulfonic acid, pH 7.5, 1 mM EDTA, 1 mM dithiothreitol and 5% glycerol, and incubated at 37 °C for 1 h. Fpg Ec (New England Biolabs) was included as the positive control. .. Products of the reactions were separated by 20% denaturing polyacrylamide gel electrophoresis (PAGE) and visualized by phosphorimaging.

    Article Title: A bipartite iron-dependent transcriptional regulation of the tryptophan salvage pathway in Chlamydia trachomatis
    Article Snippet: .. In brief, at least 3.5 μg of total RNA was incubated at 37° C with Poly(A) Polymerase in reaction buffer supplemented with ATP and murine RNase Inhibitor (New England Biolabs) for 30 min prior to heat-inactivation at 65° C for 20 min. RNA was re-isolated through an RNA clean-up filter cartridge (Ambion, ThermoFisher Scientific). ..

    Article Title: Male meiosis in Crustacea: synapsis, recombination, epigenetics and fertility in Daphnia magna
    Article Snippet: .. Following this treatment, 100 ml of reaction buffer containing 25 units of DNA polymerase I (New England BioLabs, Beverly, USA) and biotin-16-dUTP (Roche, Spain) in the nucleotide mix, was deposited onto the slide, covered with a plastic coverslip and incubated in a moist chamber for 25 min at 37 °C. .. After washing in TBE buffer (Sigma, St Louis, MO, USA), the slides were dehydrated in sequential series of ethanol baths (70, 90, and 100 % v /v ) and air-dried.

    Amplification:

    Article Title: Genetic characterization of fall armyworm infesting South Africa and India indicate recent introduction from a common source population
    Article Snippet: .. PCR amplification for all segments was performed in a 30-μl reaction mix containing 3 μl 10X manufacturer’s reaction buffer, 1 μl 10mM dNTP, 0.5 μl 20-μM primer mix, 1 μl DNA template (between 0.05–0.5 μg), 0.5 unit Taq DNA polymerase (New England Biolabs, Beverly, MA). .. Typically 96 PCR amplifications were performed at the same time using either 0.2-ml tube strips or 96 well microtiter plates.

    Article Title: Comparative molecular analyses of invasive fall armyworm in Togo reveal strong similarities to populations from the eastern United States and the Greater Antilles
    Article Snippet: .. PCR amplification for all segments was performed in a 30-μl reaction mix containing 3 μl 10X manufacturer’s reaction buffer, 1 μl 10mM dNTP, 0.5 μl 20-μM primer mix, 1 μl DNA template (between 0.05–0.5 μg), 0.5 unit Taq DNA polymerase (New England Biolabs, Beverly, MA). .. Typically 96 PCR amplifications were performed at the same time using either 0.2-ml tube strips or 96 well microtiter plates.

    Positive Control:

    Article Title: Characterization of the major formamidopyrimidine-DNA glycosylase homolog in Mycobacterium tuberculosis and its linkage to variable tandem repeats
    Article Snippet: .. The enzyme activities were assayed in a reaction buffer containing 70 mM 3-(N -morpholino) propane sulfonic acid, pH 7.5, 1 mM EDTA, 1 mM dithiothreitol and 5% glycerol, and incubated at 37 °C for 1 h. Fpg Ec (New England Biolabs) was included as the positive control. .. Products of the reactions were separated by 20% denaturing polyacrylamide gel electrophoresis (PAGE) and visualized by phosphorimaging.

    Purification:

    Article Title: Identification and Characterization of Neuronal Mitogen-activated Protein Kinase Substrates Using a Specific Phosphomotif Antibody *Identification and Characterization of Neuronal Mitogen-activated Protein Kinase Substrates Using a Specific Phosphomotif Antibody * S⃞
    Article Snippet: .. Purified protein on beads was incubated at 30 °C for 30 min in 20 μl of reaction buffer with 5 μCi of [γ-32 P]ATP, 100 μm ATP, and active purified ERK2 (100 units; New England Biolabs), JNK1 (200 ng; Millipore), or no kinase. .. Similarly PSD1 fraction (30 μg) was phosphorylated with active ERK2, JNK1, GSK-3β, CaMKII (100 units with Ca2+ /calmodulin; New England Biolabs), or cyclin-dependent kinase 2 (CDK2) (100 units with cyclin A; New England Biolabs) in the presence of 100 μm unlabeled ATP and detected by 5557 immunoblotting.

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  • 97
    New England Biolabs hf psti high fidelity
    Adapter Design, PCR amplification of fragments. 1) The ligation product of a genomic DNA fragment (black) containing a <t>PstI</t> restriction site and a <t>MspI</t> restriction site. The forward adapter (blue) binds to a PstI generated overhang. The 4–9 bp barcode for this adapter is in bold with “X”. The MspI generated overhang corresponds to the reverse Y-adapter (green). The unpaired tail of the Y-adapter is underlined. 2) During the first round of PCR only the forward primer (red) can anneal. PCR synthesis of the complementary strand proceeds to the end of the fragment synthesizing the compliment of the Y-adapter tail. 3) During the second round of PCR the reverse primer (orange) can anneal to the newly synthesized compliment of the Y-adapter tail. This PCR reaction then proceeds to fill in the compliment of the forward adapter/primer on the other end of the same fragment.
    Hf Psti High Fidelity, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hf psti high fidelity/product/New England Biolabs
    Average 97 stars, based on 3 article reviews
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    88
    New England Biolabs m sssi reaction buffer
    Cytosine deamination by <t>M.SssI</t> in vitro . pUP41 (Ap R , Kn S ) plasmid <t>DNA</t> was incubated with or without wild-type M.SssI (2-fold molar excess relative to CG sites) at 30°C for 4 h, and frequency of C-to-U deamination was determined by scoring the numbers of Kn R and Ap R transformants in E. coli ER2357 ung strain. SAM (160 µM) and 5’-amino-5’-deoxyadenosine (AA, 250 μM) were added to samples as indicated. Error bars represent standard error of the mean of at least three independent experiments (p
    M Sssi Reaction Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/m sssi reaction buffer/product/New England Biolabs
    Average 88 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
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    88
    New England Biolabs lamp reaction buffer
    Sensitivity of the reverse transcription <t>(RT)-LAMP</t> assay for detecting IBDV in bursa tissue from chickens. A — The RT-LAMP primers were used with genomic RNA for the diagnosis of IBDV infection in a 1-step RT-LAMP reaction. Lane M — 100 base pair <t>DNA</t> ladder marker; lane 1 — RT-LAMP reaction mixture containing no template; lane 2 — RT-LAMP reaction mixture containing no reverse transcriptase; lane 3 — extracted IBDV viral RNA added to the RT-LAMP reaction; lane 4 — IBDV VP2 cDNA added to the RT-LAMP reaction. B — The RT-LAMP assay and RT-polymerase chain reaction (PCR) were performed after the addition of various amounts of viral RNA. Upper panel — RT-LAMP pattern; lower panel — RT-PCR pattern. Lanes 1 to 10 — 1 ng, 100 pg, 10 pg, 1 pg, 100 fg, 10 fg, 1 fg, 0.1 fg, 0.01 fg, and 0.001 fg of IBDV total RNA, respectively. Lane NC — negative control.
    Lamp Reaction Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lamp reaction buffer/product/New England Biolabs
    Average 88 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    lamp reaction buffer - by Bioz Stars, 2020-09
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    Adapter Design, PCR amplification of fragments. 1) The ligation product of a genomic DNA fragment (black) containing a PstI restriction site and a MspI restriction site. The forward adapter (blue) binds to a PstI generated overhang. The 4–9 bp barcode for this adapter is in bold with “X”. The MspI generated overhang corresponds to the reverse Y-adapter (green). The unpaired tail of the Y-adapter is underlined. 2) During the first round of PCR only the forward primer (red) can anneal. PCR synthesis of the complementary strand proceeds to the end of the fragment synthesizing the compliment of the Y-adapter tail. 3) During the second round of PCR the reverse primer (orange) can anneal to the newly synthesized compliment of the Y-adapter tail. This PCR reaction then proceeds to fill in the compliment of the forward adapter/primer on the other end of the same fragment.

    Journal: PLoS ONE

    Article Title: Development of High-Density Genetic Maps for Barley and Wheat Using a Novel Two-Enzyme Genotyping-by-Sequencing Approach

    doi: 10.1371/journal.pone.0032253

    Figure Lengend Snippet: Adapter Design, PCR amplification of fragments. 1) The ligation product of a genomic DNA fragment (black) containing a PstI restriction site and a MspI restriction site. The forward adapter (blue) binds to a PstI generated overhang. The 4–9 bp barcode for this adapter is in bold with “X”. The MspI generated overhang corresponds to the reverse Y-adapter (green). The unpaired tail of the Y-adapter is underlined. 2) During the first round of PCR only the forward primer (red) can anneal. PCR synthesis of the complementary strand proceeds to the end of the fragment synthesizing the compliment of the Y-adapter tail. 3) During the second round of PCR the reverse primer (orange) can anneal to the newly synthesized compliment of the Y-adapter tail. This PCR reaction then proceeds to fill in the compliment of the forward adapter/primer on the other end of the same fragment.

    Article Snippet: Restriction Digest: Genomic DNA (200 ng) was digested in 20 ul reaction volume of NEB Buffer 4 with 8 U of HF-PstI (High-Fidelity) and 8 U of MspI (New England BioLabs Inc., Ipswich, MA 01938).

    Techniques: Polymerase Chain Reaction, Amplification, Ligation, Generated, Synthesized

    Cytosine deamination by M.SssI in vitro . pUP41 (Ap R , Kn S ) plasmid DNA was incubated with or without wild-type M.SssI (2-fold molar excess relative to CG sites) at 30°C for 4 h, and frequency of C-to-U deamination was determined by scoring the numbers of Kn R and Ap R transformants in E. coli ER2357 ung strain. SAM (160 µM) and 5’-amino-5’-deoxyadenosine (AA, 250 μM) were added to samples as indicated. Error bars represent standard error of the mean of at least three independent experiments (p

    Journal: PLoS ONE

    Article Title: Cytosine-to-Uracil Deamination by SssI DNA Methyltransferase

    doi: 10.1371/journal.pone.0079003

    Figure Lengend Snippet: Cytosine deamination by M.SssI in vitro . pUP41 (Ap R , Kn S ) plasmid DNA was incubated with or without wild-type M.SssI (2-fold molar excess relative to CG sites) at 30°C for 4 h, and frequency of C-to-U deamination was determined by scoring the numbers of Kn R and Ap R transformants in E. coli ER2357 ung strain. SAM (160 µM) and 5’-amino-5’-deoxyadenosine (AA, 250 μM) were added to samples as indicated. Error bars represent standard error of the mean of at least three independent experiments (p

    Article Snippet: Samples from a serial dilution of M.SssI were incubated with 0.2 - 0.5 µg plasmid or lambda phage DNA in M.SssI reaction buffer (50 mM Tris-HCl pH 8.5, 50 mM NaCl, 10 mM EDTA, 5 mM DTT containing 350 µg/ml bovine serum albumin) containing 160 μM SAM (New England Biolabs) at 30°C for one hour.

    Techniques: In Vitro, Plasmid Preparation, Incubation

    Comparison of cytosine and 5-methylcytosine deamination by M.SssI. Unmethylated and in vivo M.SssI-specifically methylated pUP41 plasmid DNA was incubated with or without M.SssI(WT) and 5’-amino-5’-deoxyadenosine as indicated using conditions described in the legend of Figure 1 . Frequency of reversion to Kn R phenotype was determined for unmethylated pUP41 in ER2357 ung (empty bars), whereas reversion frequency of methylated pUP41 was determined in DH10B ung + host (filled bars). Error bars represent standard error of the mean of at least three independent experiments (p

    Journal: PLoS ONE

    Article Title: Cytosine-to-Uracil Deamination by SssI DNA Methyltransferase

    doi: 10.1371/journal.pone.0079003

    Figure Lengend Snippet: Comparison of cytosine and 5-methylcytosine deamination by M.SssI. Unmethylated and in vivo M.SssI-specifically methylated pUP41 plasmid DNA was incubated with or without M.SssI(WT) and 5’-amino-5’-deoxyadenosine as indicated using conditions described in the legend of Figure 1 . Frequency of reversion to Kn R phenotype was determined for unmethylated pUP41 in ER2357 ung (empty bars), whereas reversion frequency of methylated pUP41 was determined in DH10B ung + host (filled bars). Error bars represent standard error of the mean of at least three independent experiments (p

    Article Snippet: Samples from a serial dilution of M.SssI were incubated with 0.2 - 0.5 µg plasmid or lambda phage DNA in M.SssI reaction buffer (50 mM Tris-HCl pH 8.5, 50 mM NaCl, 10 mM EDTA, 5 mM DTT containing 350 µg/ml bovine serum albumin) containing 160 μM SAM (New England Biolabs) at 30°C for one hour.

    Techniques: In Vivo, Methylation, Plasmid Preparation, Incubation

    Estimation of DNA MTase activity of the F17S and G19D M.SssI mutants by restriction enzyme protection assay. Lambda phage DNA was incubated with different concentrations of WT and mutant M.SssI in the presence of SAM as described in Materials and Methods. Methylation status of the DNA was subsequently tested by digestion with the methylation sensitive restriction enzyme Hin6I, and the digestion was analyzed by agarose gel electrophoresis. Lane Undig., undigested plasmid; M, molecular weight marker (GeneRuler 1 kb Plus and GeneRuler 1 kb DNA Ladders, Fermentas). M.SssI-mediated cytosine deamination in vivo was initially investigated using a two-plasmid-system, with the E. coli host containing the indicator plasmid pUP41 and one of the M.SssI-expressing plasmids pSTdC-MSssI, pSTdC-MSssI(F17S) or pSTdC-MSssI(G19D). The latter plasmids have pSC101 replicon and are compatible with pUP41. We observed elevated reversion frequency to kanamycin resistance with the Ung - host ER2357 expressing the G19D variant (not shown).

    Journal: PLoS ONE

    Article Title: Cytosine-to-Uracil Deamination by SssI DNA Methyltransferase

    doi: 10.1371/journal.pone.0079003

    Figure Lengend Snippet: Estimation of DNA MTase activity of the F17S and G19D M.SssI mutants by restriction enzyme protection assay. Lambda phage DNA was incubated with different concentrations of WT and mutant M.SssI in the presence of SAM as described in Materials and Methods. Methylation status of the DNA was subsequently tested by digestion with the methylation sensitive restriction enzyme Hin6I, and the digestion was analyzed by agarose gel electrophoresis. Lane Undig., undigested plasmid; M, molecular weight marker (GeneRuler 1 kb Plus and GeneRuler 1 kb DNA Ladders, Fermentas). M.SssI-mediated cytosine deamination in vivo was initially investigated using a two-plasmid-system, with the E. coli host containing the indicator plasmid pUP41 and one of the M.SssI-expressing plasmids pSTdC-MSssI, pSTdC-MSssI(F17S) or pSTdC-MSssI(G19D). The latter plasmids have pSC101 replicon and are compatible with pUP41. We observed elevated reversion frequency to kanamycin resistance with the Ung - host ER2357 expressing the G19D variant (not shown).

    Article Snippet: Samples from a serial dilution of M.SssI were incubated with 0.2 - 0.5 µg plasmid or lambda phage DNA in M.SssI reaction buffer (50 mM Tris-HCl pH 8.5, 50 mM NaCl, 10 mM EDTA, 5 mM DTT containing 350 µg/ml bovine serum albumin) containing 160 μM SAM (New England Biolabs) at 30°C for one hour.

    Techniques: Activity Assay, Incubation, Mutagenesis, Methylation, Agarose Gel Electrophoresis, Plasmid Preparation, Molecular Weight, Marker, In Vivo, Expressing, Variant Assay

    DNA methyltransferase activity of the F17S and G19D M.SssI mutants. ( A ) Amino acid sequence alignment between segments of M.HpaII and M.SssI. Conserved residues of the FXGXG motif are in bold. The F17S and G19D substitutions are indicated below the sequence. ( B ) Hin6I digestion of plasmids pBHNS-MSssI, pBHNS-MSssI(F17S) and pBHNS-MSssI(G19D) encoding WT or mutant M.SssI variants, respectively as indicated above the lanes. Plasmids were isolated from cultures grown for 4-6-8 h in the presence of arabinose to induce M.SssI expression. Resistance to Hin6I (recognition sequence GCGC) indicates M.SssI-specific methylation (R. Kazlauskiene, cited in REBASE [ 45 ]. Lane Unmeth., unmethylated pBHNS-MSssI(G19D) isolated from cells grown in the presence of glucose; Lane Undig., undigested pBHNS-MSssI; M, molecular weight marker (GeneRuler 1 kb DNA Ladder, Fermentas). ( C ) Effect of WT and mutant M.SssI production on growth of E. coli mcrBC and mcrBC + hosts. DH10B mcrBC contained pBHNS-MSssI, pBHNS-MSssI(F17S) or pBHNS-MSssI(G19D) as indicated. DH5α mcrBC + contained pBHNS-MSssI(F17S) or pBHNS-MSssI(G19D). Bacteria were grown in LB/Ap medium at 30°C. MTase expression was induced at time 0 by arabinose. Error bars represent standard error of the mean of three independent experiments.

    Journal: PLoS ONE

    Article Title: Cytosine-to-Uracil Deamination by SssI DNA Methyltransferase

    doi: 10.1371/journal.pone.0079003

    Figure Lengend Snippet: DNA methyltransferase activity of the F17S and G19D M.SssI mutants. ( A ) Amino acid sequence alignment between segments of M.HpaII and M.SssI. Conserved residues of the FXGXG motif are in bold. The F17S and G19D substitutions are indicated below the sequence. ( B ) Hin6I digestion of plasmids pBHNS-MSssI, pBHNS-MSssI(F17S) and pBHNS-MSssI(G19D) encoding WT or mutant M.SssI variants, respectively as indicated above the lanes. Plasmids were isolated from cultures grown for 4-6-8 h in the presence of arabinose to induce M.SssI expression. Resistance to Hin6I (recognition sequence GCGC) indicates M.SssI-specific methylation (R. Kazlauskiene, cited in REBASE [ 45 ]. Lane Unmeth., unmethylated pBHNS-MSssI(G19D) isolated from cells grown in the presence of glucose; Lane Undig., undigested pBHNS-MSssI; M, molecular weight marker (GeneRuler 1 kb DNA Ladder, Fermentas). ( C ) Effect of WT and mutant M.SssI production on growth of E. coli mcrBC and mcrBC + hosts. DH10B mcrBC contained pBHNS-MSssI, pBHNS-MSssI(F17S) or pBHNS-MSssI(G19D) as indicated. DH5α mcrBC + contained pBHNS-MSssI(F17S) or pBHNS-MSssI(G19D). Bacteria were grown in LB/Ap medium at 30°C. MTase expression was induced at time 0 by arabinose. Error bars represent standard error of the mean of three independent experiments.

    Article Snippet: Samples from a serial dilution of M.SssI were incubated with 0.2 - 0.5 µg plasmid or lambda phage DNA in M.SssI reaction buffer (50 mM Tris-HCl pH 8.5, 50 mM NaCl, 10 mM EDTA, 5 mM DTT containing 350 µg/ml bovine serum albumin) containing 160 μM SAM (New England Biolabs) at 30°C for one hour.

    Techniques: Activity Assay, Sequencing, Mutagenesis, Isolation, Expressing, Methylation, Molecular Weight, Marker

    Sensitivity of the reverse transcription (RT)-LAMP assay for detecting IBDV in bursa tissue from chickens. A — The RT-LAMP primers were used with genomic RNA for the diagnosis of IBDV infection in a 1-step RT-LAMP reaction. Lane M — 100 base pair DNA ladder marker; lane 1 — RT-LAMP reaction mixture containing no template; lane 2 — RT-LAMP reaction mixture containing no reverse transcriptase; lane 3 — extracted IBDV viral RNA added to the RT-LAMP reaction; lane 4 — IBDV VP2 cDNA added to the RT-LAMP reaction. B — The RT-LAMP assay and RT-polymerase chain reaction (PCR) were performed after the addition of various amounts of viral RNA. Upper panel — RT-LAMP pattern; lower panel — RT-PCR pattern. Lanes 1 to 10 — 1 ng, 100 pg, 10 pg, 1 pg, 100 fg, 10 fg, 1 fg, 0.1 fg, 0.01 fg, and 0.001 fg of IBDV total RNA, respectively. Lane NC — negative control.

    Journal: Canadian Journal of Veterinary Research

    Article Title: One-step reverse-transcription loop-mediated isothermal amplification for detection of infectious bursal disease virus

    doi:

    Figure Lengend Snippet: Sensitivity of the reverse transcription (RT)-LAMP assay for detecting IBDV in bursa tissue from chickens. A — The RT-LAMP primers were used with genomic RNA for the diagnosis of IBDV infection in a 1-step RT-LAMP reaction. Lane M — 100 base pair DNA ladder marker; lane 1 — RT-LAMP reaction mixture containing no template; lane 2 — RT-LAMP reaction mixture containing no reverse transcriptase; lane 3 — extracted IBDV viral RNA added to the RT-LAMP reaction; lane 4 — IBDV VP2 cDNA added to the RT-LAMP reaction. B — The RT-LAMP assay and RT-polymerase chain reaction (PCR) were performed after the addition of various amounts of viral RNA. Upper panel — RT-LAMP pattern; lower panel — RT-PCR pattern. Lanes 1 to 10 — 1 ng, 100 pg, 10 pg, 1 pg, 100 fg, 10 fg, 1 fg, 0.1 fg, 0.01 fg, and 0.001 fg of IBDV total RNA, respectively. Lane NC — negative control.

    Article Snippet: The reaction mixture contained 12.5 μL of 2× LAMP reaction buffer , 8 U of Bst DNA polymerase (New England Biolabs, Frankfurt, Germany), 10 U of AMV RTase (Invitrogen), 10 μM of each of the F3 and B3 primers, 10 μM of each of the FIP and BIP primers, 10 μM of 2 M betaine, and 2 μL of the viral RNA.

    Techniques: RT Lamp Assay, Infection, Marker, Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Negative Control

    Specificity of the loop-mediated isothermal amplification (LAMP) assay for detecting infectious bursal disease virus (IBDV). The templates were IBDV VP2 complementary DNA (cDNA), the VP2 gene of chicken anemia virus (CAV), and the H6 gene of avian influenza virus (AIV) H6N1. Only for IBDV were LAMP products visualized with (A) 1.6% agarose gel electrophoresis and (B) staining with SYBR Green I and ultraviolet excitation. Lane C — negative control.

    Journal: Canadian Journal of Veterinary Research

    Article Title: One-step reverse-transcription loop-mediated isothermal amplification for detection of infectious bursal disease virus

    doi:

    Figure Lengend Snippet: Specificity of the loop-mediated isothermal amplification (LAMP) assay for detecting infectious bursal disease virus (IBDV). The templates were IBDV VP2 complementary DNA (cDNA), the VP2 gene of chicken anemia virus (CAV), and the H6 gene of avian influenza virus (AIV) H6N1. Only for IBDV were LAMP products visualized with (A) 1.6% agarose gel electrophoresis and (B) staining with SYBR Green I and ultraviolet excitation. Lane C — negative control.

    Article Snippet: The reaction mixture contained 12.5 μL of 2× LAMP reaction buffer , 8 U of Bst DNA polymerase (New England Biolabs, Frankfurt, Germany), 10 U of AMV RTase (Invitrogen), 10 μM of each of the F3 and B3 primers, 10 μM of each of the FIP and BIP primers, 10 μM of 2 M betaine, and 2 μL of the viral RNA.

    Techniques: Amplification, Lamp Assay, Agarose Gel Electrophoresis, Staining, SYBR Green Assay, Negative Control

    Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. meningitidis with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.

    Journal: Biosensors & bioelectronics

    Article Title: Fully 3D Printed Integrated Reactor Array for Point-of-Care Molecular Diagnostics

    doi: 10.1016/j.bios.2018.03.009

    Figure Lengend Snippet: Colorimetric and fluorescence based detection for NAATs in 3D printed reactor array. A) Representative photographs of colorimetric LAMP assay for detection of N. meningitidis with 0, 50, 500 and 5000 CFU/reaction on the same chip, alongside LAMP fluorescence based image at given time interval. B) LAMP amplification curves for P. falciparum with 0, 0.1 1, 10, 100, 1000 pg per reaction. C) Calibration curve for P. falciparum as function of log target concentration, n=3. D) LAMP amplification curves for N. meningitidis with 0, 50, 500, 5000 CFU per reaction. E) Calibration curve for N. meningitidis as function of log target concentration, n=3. WarmStart ® LAMP master mix was used.

    Article Snippet: Bovine serum albumin (BSA), Poly (ethylene glycol) 8000 (PEG), poly(vinyl alcohol) (PVA) are from Sigma-Aldrich, RT-PCR grade water from Ambion, Inc., intercalating Eva Green fluorescent dye from Biotium, Isothermal Master Mix used in LAMP reaction buffer from ISO-001nd, OptiGene, Horsham, UK, WarmStart® colorimetric LAMP 2X master mix from New England Biolabs Inc. N. meningitidis (ATCC 13098) from American Type Culture Collection (ATCC, Rockville, MD).

    Techniques: Fluorescence, Lamp Assay, Chromatin Immunoprecipitation, Amplification, Concentration Assay

    LAMP amplification curves of serial dilutions of P. falciparum gDNA in PBS samples with or without static coating in the 3D printed amplification reactors. A) no coating, B) BSA coating, C) PEG coating and D) PVA coating (n=3). Optigene ® LAMP Isothermal Master Mix was used. Note: rxn = reaction

    Journal: Biosensors & bioelectronics

    Article Title: Fully 3D Printed Integrated Reactor Array for Point-of-Care Molecular Diagnostics

    doi: 10.1016/j.bios.2018.03.009

    Figure Lengend Snippet: LAMP amplification curves of serial dilutions of P. falciparum gDNA in PBS samples with or without static coating in the 3D printed amplification reactors. A) no coating, B) BSA coating, C) PEG coating and D) PVA coating (n=3). Optigene ® LAMP Isothermal Master Mix was used. Note: rxn = reaction

    Article Snippet: Bovine serum albumin (BSA), Poly (ethylene glycol) 8000 (PEG), poly(vinyl alcohol) (PVA) are from Sigma-Aldrich, RT-PCR grade water from Ambion, Inc., intercalating Eva Green fluorescent dye from Biotium, Isothermal Master Mix used in LAMP reaction buffer from ISO-001nd, OptiGene, Horsham, UK, WarmStart® colorimetric LAMP 2X master mix from New England Biolabs Inc. N. meningitidis (ATCC 13098) from American Type Culture Collection (ATCC, Rockville, MD).

    Techniques: Amplification