taq  (New England Biolabs)


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  • 86
    Name:
    Standard Taq Reaction Buffer Pack
    Description:
    Standard Taq Reaction Buffer Pack 6 0 ml
    Catalog Number:
    B9014S
    Price:
    22
    Category:
    Buffers
    Size:
    6 0 ml
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    Structured Review

    New England Biolabs taq
    Standard Taq Reaction Buffer Pack
    Standard Taq Reaction Buffer Pack 6 0 ml
    https://www.bioz.com/result/taq/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    taq - by Bioz Stars, 2021-05
    86/100 stars

    Images

    1) Product Images from "Determining the One, Two, Three, or Four Long and Short Loci of Human Complement C4 in a Major Histocompatibility Complex Haplotype Encoding C4A or C4B Proteins"

    Article Title: Determining the One, Two, Three, or Four Long and Short Loci of Human Complement C4 in a Major Histocompatibility Complex Haplotype Encoding C4A or C4B Proteins

    Journal: American Journal of Human Genetics

    doi:

    Defining the RCCX modular structure using different genomic RFLP Southern blot analyses. A, Restriction patterns of Taq I RFLP for the five selected individuals on simultaneous hybridization of probes specific for 3′ RP, CYP21, and 3′ TNX. B, Psh AI RFLP hybridized to a 3′ RP probe. C, Bam HI RFLP hybridized to a 3′ TNX probe. D, Phenotyping of C4A and C4B proteins by immunofixation of EDTA-blood plasma resolved by HVAGE. E, Immunoblot analysis of Rg1 and Ch1 antigenic determinants in plasma C4 proteins after HVAGE. F, Genomic Southern blot analysis of C4A and C4B genes by Psh AI-RFLP (using Probe D) and by Psh AI- Pvu II RFLP (using probe 22–25). G, HLA class I and class II alleles of the five subjects.
    Figure Legend Snippet: Defining the RCCX modular structure using different genomic RFLP Southern blot analyses. A, Restriction patterns of Taq I RFLP for the five selected individuals on simultaneous hybridization of probes specific for 3′ RP, CYP21, and 3′ TNX. B, Psh AI RFLP hybridized to a 3′ RP probe. C, Bam HI RFLP hybridized to a 3′ TNX probe. D, Phenotyping of C4A and C4B proteins by immunofixation of EDTA-blood plasma resolved by HVAGE. E, Immunoblot analysis of Rg1 and Ch1 antigenic determinants in plasma C4 proteins after HVAGE. F, Genomic Southern blot analysis of C4A and C4B genes by Psh AI-RFLP (using Probe D) and by Psh AI- Pvu II RFLP (using probe 22–25). G, HLA class I and class II alleles of the five subjects.

    Techniques Used: Southern Blot, Hybridization

    2) Product Images from "A Recurrent Stop-Codon Mutation in Succinate Dehydrogenase Subunit B Gene in Normal Peripheral Blood and Childhood T-Cell Acute Leukemia"

    Article Title: A Recurrent Stop-Codon Mutation in Succinate Dehydrogenase Subunit B Gene in Normal Peripheral Blood and Childhood T-Cell Acute Leukemia

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0000436

    SDHB mutations in genomic DNAs (gDNAs). A. Taq I RE digestion of PCR-amplified exon 2 shows no evidence of undigested mutant DNAs at 260 bp size (pointed by an arrow head). Taq I RE digestion products of the wild-type DNA co-migrate at sizes of 128 and 132 bps (denoted by a star). B. Gel electrophoresis of Taq I RE-digested (mutation) enrichment PCR ( e PCR) products shows variable amounts of mutant gDNAs at 233 bp size (pointed by an arrow head), which are confirmed to be primarily composed of c.136C > T by sequencing ( Table 1 ). Taq I digestion of wild-type e PCR products gives two bands at sizes, 101 and 132 bp (shown by a star). C. e PCR analyses of leukemic cell lines demonstrate the presence of mutant gDNA in the MOLT-4 cell line (pointed by an arrow head) but not in other leukemic cell lines in this experiment. D. Direct sequence analysis of a gDNA e PCR product shows selective enrichment of the R46X mutation. E, F. Sequence chromatograms of the heterozygous DNA mutations detected in two T-ALL cell lines are shown.
    Figure Legend Snippet: SDHB mutations in genomic DNAs (gDNAs). A. Taq I RE digestion of PCR-amplified exon 2 shows no evidence of undigested mutant DNAs at 260 bp size (pointed by an arrow head). Taq I RE digestion products of the wild-type DNA co-migrate at sizes of 128 and 132 bps (denoted by a star). B. Gel electrophoresis of Taq I RE-digested (mutation) enrichment PCR ( e PCR) products shows variable amounts of mutant gDNAs at 233 bp size (pointed by an arrow head), which are confirmed to be primarily composed of c.136C > T by sequencing ( Table 1 ). Taq I digestion of wild-type e PCR products gives two bands at sizes, 101 and 132 bp (shown by a star). C. e PCR analyses of leukemic cell lines demonstrate the presence of mutant gDNA in the MOLT-4 cell line (pointed by an arrow head) but not in other leukemic cell lines in this experiment. D. Direct sequence analysis of a gDNA e PCR product shows selective enrichment of the R46X mutation. E, F. Sequence chromatograms of the heterozygous DNA mutations detected in two T-ALL cell lines are shown.

    Techniques Used: Polymerase Chain Reaction, Amplification, Mutagenesis, Nucleic Acid Electrophoresis, Sequencing

    Related Articles

    Polymerase Chain Reaction:

    Article Title: Breaking the speed limit with multimode fast scanning of DNA by Endonuclease V
    Article Snippet: This DNA substrate was prepared by PCR amplification of λ-DNA using primers modified with 5′ biotin (5′-bio-ACTTCGCCTTCTTCCCATTT-3′) and 5′ digoxigenin (5′-dig-ATCTCGCTTTCCACTCCAGA-3′) (Eurofins MWG/Operon). .. The PCR reaction was performed in a 1× LongAmp Taq Reaction Buffer, with a final volume of 50 μl, 300 µM of each dNTP, 0.4 µM of each primer, 2 units LongAmp Taq DNA polymerase (New England Biolabs) and 0.1 ng µl−1 of λ-DNA template. .. The PCR included an initial denaturation step at 94 °C for 3 min, 35 cycles of denaturation (94 °C for 15 s), annealing (60 °C for 60 s) and primer extension (65 °C for 16 min), followed by a final extension step at 65 °C for 10 min.

    Generated:

    Article Title: In Vitro Fluorogenic Real-time Assay of the Repair of Oxidative DNA Damage
    Article Snippet: The initial velocity was determined by measuring the rate of fluorescence increase at 460 nm over the first 10 minutes of the reaction, during which the fluorescence increase was linear. .. Michaelis Menten curves for Fpg were generated by incubating probe OGR1 (0.03-0.3 μM) with 6 nM Fpg in Fpg reaction buffer and 100 μg/mL BSA at 37 °C and measuring the initial velocity at 37 °C. .. The initial velocity was determined by measuring the rate of fluorescence increase at 460 nm over the first 6 minutes of the reaction, during which the fluorescence increase was linear.

    Article Title: Rolling Circle DNA Synthesis: Small Circular Oligonucleotides as Efficient Templates for DNA Polymerases
    Article Snippet: The reaction mixture was the same as the standard conditions described above except with 1 mM each of dATP, dCTP, and dGTP, 0.165 mM of α-[32 P]-dTTP (specific activity: 3000 Ci/mmol). .. Reaction products generated after 3 h of synthesis were precipitated by adding two volumes of precipitation buffer (50 mM KCl in ethanol) and were then resuspended in 20 µL of Taq Iα reaction buffer: 10 mM NaCl, 1 mM MgCl2 , 100 mM Tris•HCl (pH 8.4), 10 mg/mL BSA, with 5 u/µL Taq Iα (New England Biolabs). ..

    Incubation:

    Article Title: Genotyping of Epidemic Methicillin-Resistant Staphylococcus aureus Phage Type 15 Isolates by Fluorescent Amplified-Fragment Length Polymorphism Analysis
    Article Snippet: .. The enzymes Apa I and Taq I were used to digest DNA as follows: approximately 500 ng of DNA was incubated with 4 U of Apa I (NEB), 1× buffer 4 (NEB), 1× bovine serum albumin (NEB), and 0.5 mg of DNase-free RNase A ml−1 in a final volume of 20 μl at 25°C for 1 h. Five units of Taq I (NEB) was subsequently added to each reaction mixture, and the mixtures were incubated for a further 1 h at 65°C. .. A 20-μl solution containing 4 pmol of Apa I adaptors (MWG-Biotech UK Ltd., Milton Keynes, United Kingdom), 40 pmol of Taq I adaptors (MWG-Biotech), 1× T4 ligase buffer (NEB), and 40 U of T4 DNA ligase (NEB) was added to 20 μl of double-digested DNA.

    Article Title: Potassium Dependent Regulation of Astrocyte Water Permeability Is Mediated by cAMP Signaling
    Article Snippet: In Vitro Phosphorylation In vitro phosphorylation was performed as described previously . .. Briefly, purified glutathione S-transferase (GST) fusion proteins, GST-APQ4 S111 or GST-APQ4 A111, were incubated with cAMP-dependent Protein Kinase (PKA), catalytic subunit (New England Biolabs) in PKA reaction buffer (New England Biolabs) containing 200 µM ATP, γ-32 pATP (0.2 µCi/lL), and Phosphatase Inhibitor Cocktail 2 (Sigma). ..

    Transformation Assay:

    Article Title: Primer Extension Mutagenesis Powered by Selective Rolling Circle Amplification
    Article Snippet: The oligo was phosphorylated, hybridized to ss(U)DNA and extended as described in Kunkel mutagenesis for CDR-H1 loop. .. The obtained heteroduplex was either directly transformed to E. coli XL-1 Blue or subjected to an UDG treatment, in which, 1 µg Kunkel heteroduplex was treated in 1× UDG reaction buffer (NEB, Ipswich, USA) containing 20 mM Tris-HCl (pH 8.0), 1 mM EDTA, 1 mM DTT and 10 U UDG (NEB, Ipswich, USA) for 1 h at 37°C in a total volume of 25 µl. .. A control sample without UDG treatment was incubated in the same conditions as the UDG treated sample with the exception that the UDG was replaced with H2 O.

    Purification:

    Article Title: Potassium Dependent Regulation of Astrocyte Water Permeability Is Mediated by cAMP Signaling
    Article Snippet: In Vitro Phosphorylation In vitro phosphorylation was performed as described previously . .. Briefly, purified glutathione S-transferase (GST) fusion proteins, GST-APQ4 S111 or GST-APQ4 A111, were incubated with cAMP-dependent Protein Kinase (PKA), catalytic subunit (New England Biolabs) in PKA reaction buffer (New England Biolabs) containing 200 µM ATP, γ-32 pATP (0.2 µCi/lL), and Phosphatase Inhibitor Cocktail 2 (Sigma). ..

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  • 99
    New England Biolabs dna buffer
    DNase I footprinting assays monitoring PIC assembly and structural isomerization. ( A ) Early steps of PIC assembly. Specified GTFs were incubated with the end-labeled <t>DNA</t> template, followed by DNase I digestion. Blue bracket highlights the <t>TFIID</t> footprint. The numbers are relative to the transcription start site (+1). ( B ) Arresting of the conformational isomerization. Top, the scheme. The inhibitor used here was SnOCl 2 ·pyridine. Lanes 1 and 9 are 10 bp DNA ladder. Lanes 2 and 10 are digestion of naked DNA. The lower case ‘d’ and ‘f’ in lanes #11–14 reflect the use of less TFIID and TFIIF (together with the omission of spermidine and carrier nucleic acid in the reaction—see ‘Material and methods’ for detail). Letter abbreviations are explained in Figure 6 legend. DOI: http://dx.doi.org/10.7554/eLife.07777.016
    Dna Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna buffer/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
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    99
    New England Biolabs taq polymerase
    Amplification results of three target genes using <t>Taq</t> polymerase. (A) Attempted amplification of the three <t>DNA</t> targets ( mpb83 , Mb012, and LMHCC_RS00060 ) sequences by Taq in the presence of DMSO using 3St, 2St and TD protocols; mpb83 and LMHCC_RS00060 successfully showed amplicons using the three protocols. Mb012 failed to show an amplicon. (B) Attempts of amplification of the three DNA target sequences in the absence of DMSO using 3St, 2St and TD protocols showing no amplicon. mpb83 with added enhancer in 3St PCR is used as a positive control. Mb0129 with no enhancer in 3St PCR is used as a negative control.
    Taq Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taq polymerase/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    taq polymerase - by Bioz Stars, 2021-05
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    99
    New England Biolabs crimson longamp taq polymerase
    Derivative dissociation plots produced by different reagents using one microliter of lysate produced by homogenizing one H. armigera leg with 24 H. zea legs in 625 μl of modified squish buffer. (A) Amplification reactions made with KAPA SYBR Fast Master Mix (purple) and NEB <t>LongAmp</t> <t>Taq</t> polymerase and buffer with 2.5 mM MgCl2 (Red). Peaks representing 18S rRNA control amplicon and species-specific amplicons of H. armigera and H. zea are present in all reactions, but melting temperatures were different in two reagents. (B) An enlarged view of the derivative dissociation plot produced by amplification reactions with NEB LongAmp Taq polymerase and buffer. Although variations in dissociation plots between samples were observed, the 3-peak dissociation plot was clearly identifiable.
    Crimson Longamp Taq Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/crimson longamp taq polymerase/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    DNase I footprinting assays monitoring PIC assembly and structural isomerization. ( A ) Early steps of PIC assembly. Specified GTFs were incubated with the end-labeled DNA template, followed by DNase I digestion. Blue bracket highlights the TFIID footprint. The numbers are relative to the transcription start site (+1). ( B ) Arresting of the conformational isomerization. Top, the scheme. The inhibitor used here was SnOCl 2 ·pyridine. Lanes 1 and 9 are 10 bp DNA ladder. Lanes 2 and 10 are digestion of naked DNA. The lower case ‘d’ and ‘f’ in lanes #11–14 reflect the use of less TFIID and TFIIF (together with the omission of spermidine and carrier nucleic acid in the reaction—see ‘Material and methods’ for detail). Letter abbreviations are explained in Figure 6 legend. DOI: http://dx.doi.org/10.7554/eLife.07777.016

    Journal: eLife

    Article Title: Chemical perturbation of an intrinsically disordered region of TFIID distinguishes two modes of transcription initiation

    doi: 10.7554/eLife.07777

    Figure Lengend Snippet: DNase I footprinting assays monitoring PIC assembly and structural isomerization. ( A ) Early steps of PIC assembly. Specified GTFs were incubated with the end-labeled DNA template, followed by DNase I digestion. Blue bracket highlights the TFIID footprint. The numbers are relative to the transcription start site (+1). ( B ) Arresting of the conformational isomerization. Top, the scheme. The inhibitor used here was SnOCl 2 ·pyridine. Lanes 1 and 9 are 10 bp DNA ladder. Lanes 2 and 10 are digestion of naked DNA. The lower case ‘d’ and ‘f’ in lanes #11–14 reflect the use of less TFIID and TFIIF (together with the omission of spermidine and carrier nucleic acid in the reaction—see ‘Material and methods’ for detail). Letter abbreviations are explained in Figure 6 legend. DOI: http://dx.doi.org/10.7554/eLife.07777.016

    Article Snippet: The experiment in lanes 9–14 was performed slightly differently in that: (1) the TFIID amount was 10 ng and TFIIF was 2 ng; (2) the DNA buffer was replaced with (10 mM Tris pH 8.0, and 0.1 mM EDTA), and the carrier nucleic acids were eliminated; and (3) 2 µl 20 mU/µl DNase I (NEB) was used for the digestion.

    Techniques: Footprinting, Incubation, Labeling

    A model of inhibition that mechanistically distinguishes the two modes of transcription initiation. ( A ) Initially, TFIID forms multiple contacts with an extended promoter DNA region that is stabilized by TFIIA and TFIIB. TFIIA doesn't affect transcription at this promoter with purified factors, but it does facilitate the TATA box protection by TFIID alone ( Cianfrocco et al., 2013 ) or by TFIID together with TFIIB (ZZ and RT unpublished). We propose a critical isomerization step during de novo PIC assembly involving a TFIID conformational change (i.e., release of at least part of the promoter DNA, illustrated by the change in the shape of TFIID) to allow entry and engagement of Pol II. Once Pol II becomes engaged and further stabilized by other factors (TFIIE, TFIIF, etc) transcription can proceed. ( B ) The inhibitor, by binding and interfering with the TAF2 IDR, arrests TFIID isomerization and Pol II engagement, thus, blocking the assembly of a functional PIC. DNase I footprint assay reveals that Pol II molecules can still partially interact with the downstream portion of promoter DNA in the presence of the inhibitor. ( C ) Once the first round of Pol II engagement is accomplished and isomerization has occurred, the PIC intermediate establishes a state resistant to inhibition. After Pol II enters the elongation phase, TFIID remains at the isomerized state as part of a reinitiation scaffold. This reinitiation complex bypasses the initial stages of de novo PIC assembly where TFIID contacts an extended DNA region and thus is resistant to the inhibition by the tin(IV) oxochloride cluster. In addition, this shortcut may be a mechanism for the reinitiation scaffold to facilitate reloading of more Pol II molecules. DOI: http://dx.doi.org/10.7554/eLife.07777.019

    Journal: eLife

    Article Title: Chemical perturbation of an intrinsically disordered region of TFIID distinguishes two modes of transcription initiation

    doi: 10.7554/eLife.07777

    Figure Lengend Snippet: A model of inhibition that mechanistically distinguishes the two modes of transcription initiation. ( A ) Initially, TFIID forms multiple contacts with an extended promoter DNA region that is stabilized by TFIIA and TFIIB. TFIIA doesn't affect transcription at this promoter with purified factors, but it does facilitate the TATA box protection by TFIID alone ( Cianfrocco et al., 2013 ) or by TFIID together with TFIIB (ZZ and RT unpublished). We propose a critical isomerization step during de novo PIC assembly involving a TFIID conformational change (i.e., release of at least part of the promoter DNA, illustrated by the change in the shape of TFIID) to allow entry and engagement of Pol II. Once Pol II becomes engaged and further stabilized by other factors (TFIIE, TFIIF, etc) transcription can proceed. ( B ) The inhibitor, by binding and interfering with the TAF2 IDR, arrests TFIID isomerization and Pol II engagement, thus, blocking the assembly of a functional PIC. DNase I footprint assay reveals that Pol II molecules can still partially interact with the downstream portion of promoter DNA in the presence of the inhibitor. ( C ) Once the first round of Pol II engagement is accomplished and isomerization has occurred, the PIC intermediate establishes a state resistant to inhibition. After Pol II enters the elongation phase, TFIID remains at the isomerized state as part of a reinitiation scaffold. This reinitiation complex bypasses the initial stages of de novo PIC assembly where TFIID contacts an extended DNA region and thus is resistant to the inhibition by the tin(IV) oxochloride cluster. In addition, this shortcut may be a mechanism for the reinitiation scaffold to facilitate reloading of more Pol II molecules. DOI: http://dx.doi.org/10.7554/eLife.07777.019

    Article Snippet: The experiment in lanes 9–14 was performed slightly differently in that: (1) the TFIID amount was 10 ng and TFIIF was 2 ng; (2) the DNA buffer was replaced with (10 mM Tris pH 8.0, and 0.1 mM EDTA), and the carrier nucleic acids were eliminated; and (3) 2 µl 20 mU/µl DNase I (NEB) was used for the digestion.

    Techniques: Inhibition, Purification, Binding Assay, Blocking Assay, Functional Assay

    Amplification results of three target genes using Taq polymerase. (A) Attempted amplification of the three DNA targets ( mpb83 , Mb012, and LMHCC_RS00060 ) sequences by Taq in the presence of DMSO using 3St, 2St and TD protocols; mpb83 and LMHCC_RS00060 successfully showed amplicons using the three protocols. Mb012 failed to show an amplicon. (B) Attempts of amplification of the three DNA target sequences in the absence of DMSO using 3St, 2St and TD protocols showing no amplicon. mpb83 with added enhancer in 3St PCR is used as a positive control. Mb0129 with no enhancer in 3St PCR is used as a negative control.

    Journal: bioRxiv

    Article Title: PCR procedures to amplify GC-rich DNA sequences of Mycobacterium bovis

    doi: 10.1101/2020.02.18.953695

    Figure Lengend Snippet: Amplification results of three target genes using Taq polymerase. (A) Attempted amplification of the three DNA targets ( mpb83 , Mb012, and LMHCC_RS00060 ) sequences by Taq in the presence of DMSO using 3St, 2St and TD protocols; mpb83 and LMHCC_RS00060 successfully showed amplicons using the three protocols. Mb012 failed to show an amplicon. (B) Attempts of amplification of the three DNA target sequences in the absence of DMSO using 3St, 2St and TD protocols showing no amplicon. mpb83 with added enhancer in 3St PCR is used as a positive control. Mb0129 with no enhancer in 3St PCR is used as a negative control.

    Article Snippet: Unsuccessful attempts were faced with Taq polymerase, OneTaq DNA Polymerase (NEB, M0480S), Platinum™ Pfx DNA Polymerase (Invitrogen, 11708039), Expand Long Template PCR System (an enzyme mix that contains thermostable Taq DNA Polymerase and a thermostable DNA polymerase with proofreading activity, Roche, 11681834001).

    Techniques: Amplification, Polymerase Chain Reaction, Positive Control, Negative Control

    PEAR reactions with complete and incomplete components. Target (X) and probe (X′R′X′R′X′) concentrations were at 1nM and 100 nM respectively. For PspGI, H and L stand for high (0.4 U/µL) and low (0.1 U/µL) concentrations respectively. Lane M: Invitrogen Trackit™ 10 bp DNA ladder; Lane 1–2: complete PEAR reactions containing Taq DNA polymerase, PspGI, the target and the probe. The lower band (shown by an arrow) represents the 20-bp duplex monomers, and the upper bands represent tandem repeats; Lane 3–9: incomplete PEAR reactions lacking one or both enzymes or the target. No product band was observed. The bands represent probe self-dimerization formed by intermolecular interactions.

    Journal: PLoS ONE

    Article Title: Polymerase-Endonuclease Amplification Reaction (PEAR) for Large-Scale Enzymatic Production of Antisense Oligonucleotides

    doi: 10.1371/journal.pone.0008430

    Figure Lengend Snippet: PEAR reactions with complete and incomplete components. Target (X) and probe (X′R′X′R′X′) concentrations were at 1nM and 100 nM respectively. For PspGI, H and L stand for high (0.4 U/µL) and low (0.1 U/µL) concentrations respectively. Lane M: Invitrogen Trackit™ 10 bp DNA ladder; Lane 1–2: complete PEAR reactions containing Taq DNA polymerase, PspGI, the target and the probe. The lower band (shown by an arrow) represents the 20-bp duplex monomers, and the upper bands represent tandem repeats; Lane 3–9: incomplete PEAR reactions lacking one or both enzymes or the target. No product band was observed. The bands represent probe self-dimerization formed by intermolecular interactions.

    Article Snippet: Alternatively, in order to remove BSA, Taq DNA polymerases and excess dNTPs, PEAR products were phenol extracted, ethanol precipitated, washed three times with 75% ethanol, dried and resuspended in ddH2 O, and double digested by the addition of 1 volume of cleavage mixture containing 1× NEBuffer 4, and 1.0 u/µl each of Hpy99I and PspGI.

    Techniques:

    Schematic description of PEAR. Sense and antisense strands are represented by solid and dashed lines respectively, the 3′ ends are indicated by arrows and the restriction sites for PspGI are indicated by solid diamonds. When a target oligonucleotide ( X ) binds to a probe in the upstream, it is elongated by the Taq DNA polymerase, and a full-duplex oligonucleotide containing tandem repeats is produced. If the repeats are cleaved by PspGI, short duplex oligos ( X/X′ ) are released; and when they are not cleaved, the number of tandem repeats increases by slipping and elongation.

    Journal: PLoS ONE

    Article Title: Polymerase-Endonuclease Amplification Reaction (PEAR) for Large-Scale Enzymatic Production of Antisense Oligonucleotides

    doi: 10.1371/journal.pone.0008430

    Figure Lengend Snippet: Schematic description of PEAR. Sense and antisense strands are represented by solid and dashed lines respectively, the 3′ ends are indicated by arrows and the restriction sites for PspGI are indicated by solid diamonds. When a target oligonucleotide ( X ) binds to a probe in the upstream, it is elongated by the Taq DNA polymerase, and a full-duplex oligonucleotide containing tandem repeats is produced. If the repeats are cleaved by PspGI, short duplex oligos ( X/X′ ) are released; and when they are not cleaved, the number of tandem repeats increases by slipping and elongation.

    Article Snippet: Alternatively, in order to remove BSA, Taq DNA polymerases and excess dNTPs, PEAR products were phenol extracted, ethanol precipitated, washed three times with 75% ethanol, dried and resuspended in ddH2 O, and double digested by the addition of 1 volume of cleavage mixture containing 1× NEBuffer 4, and 1.0 u/µl each of Hpy99I and PspGI.

    Techniques: Produced

    Derivative dissociation plots produced by different reagents using one microliter of lysate produced by homogenizing one H. armigera leg with 24 H. zea legs in 625 μl of modified squish buffer. (A) Amplification reactions made with KAPA SYBR Fast Master Mix (purple) and NEB LongAmp Taq polymerase and buffer with 2.5 mM MgCl2 (Red). Peaks representing 18S rRNA control amplicon and species-specific amplicons of H. armigera and H. zea are present in all reactions, but melting temperatures were different in two reagents. (B) An enlarged view of the derivative dissociation plot produced by amplification reactions with NEB LongAmp Taq polymerase and buffer. Although variations in dissociation plots between samples were observed, the 3-peak dissociation plot was clearly identifiable.

    Journal: Journal of Insect Science

    Article Title: Rapid Identification of Helicoverpa armigera and Helicoverpa zea (Lepidoptera: Noctuidae) Using Ribosomal RNA Internal Transcribed Spacer 1

    doi: 10.1093/jisesa/iev137

    Figure Lengend Snippet: Derivative dissociation plots produced by different reagents using one microliter of lysate produced by homogenizing one H. armigera leg with 24 H. zea legs in 625 μl of modified squish buffer. (A) Amplification reactions made with KAPA SYBR Fast Master Mix (purple) and NEB LongAmp Taq polymerase and buffer with 2.5 mM MgCl2 (Red). Peaks representing 18S rRNA control amplicon and species-specific amplicons of H. armigera and H. zea are present in all reactions, but melting temperatures were different in two reagents. (B) An enlarged view of the derivative dissociation plot produced by amplification reactions with NEB LongAmp Taq polymerase and buffer. Although variations in dissociation plots between samples were observed, the 3-peak dissociation plot was clearly identifiable.

    Article Snippet: Initial tests of species-specific fragment amplification were carried out using Crimson LongAmp Taq polymerase with a 45 s initial denaturation step at 95°C followed by 35 cycles of 15 s denaturation at 95°C, 10 s annealing at 60°C, and 30 s extension at 72°C.

    Techniques: Produced, Modification, Amplification