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    New England Biolabs q5 highfidelity dna polymerase
    Olig2.5 inhibited archaeal family B <t>DNA</t> polymerases. An E. coli DH5α colony harbouring the 2.6 kb upstream sequence of Olig2 in pGL3-Basic vector was cultured and 1 μl of the culture was directly used as template for PCR. Olig2F and OligTSSR were used as primers to amplify the 2 kb fragment. Olig2.5 was added to see its inhibition to various DNAP. DNAP: DNA polymerase. LA: LA Taq DNAP. Q5: Q5 High-Fidelity DNAP. PS: PrimeSTAR HS DNAP. GXL: PrimeSTAR GXL DNAP. Phusion: Phusion High-Fidelity DNAP. Cobuddy: Cobuddy Super Fidelity DNAP. KOD: KOD DNAP.
    Q5 Highfidelity Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2358 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 highfidelity dna polymerase/product/New England Biolabs
    Average 99 stars, based on 2358 article reviews
    Price from $9.99 to $1999.99
    q5 highfidelity dna polymerase - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    99
    New England Biolabs q5 high fidelity
    Olig2.5 inhibited archaeal family B <t>DNA</t> polymerases. An E. coli DH5α colony harbouring the 2.6 kb upstream sequence of Olig2 in pGL3-Basic vector was cultured and 1 μl of the culture was directly used as template for PCR. Olig2F and OligTSSR were used as primers to amplify the 2 kb fragment. Olig2.5 was added to see its inhibition to various DNAP. DNAP: DNA polymerase. LA: LA Taq DNAP. Q5: Q5 High-Fidelity DNAP. PS: PrimeSTAR HS DNAP. GXL: PrimeSTAR GXL DNAP. Phusion: Phusion High-Fidelity DNAP. Cobuddy: Cobuddy Super Fidelity DNAP. KOD: KOD DNAP.
    Q5 High Fidelity, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 499 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 high fidelity/product/New England Biolabs
    Average 99 stars, based on 499 article reviews
    Price from $9.99 to $1999.99
    q5 high fidelity - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

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    Olig2.5 inhibited archaeal family B DNA polymerases. An E. coli DH5α colony harbouring the 2.6 kb upstream sequence of Olig2 in pGL3-Basic vector was cultured and 1 μl of the culture was directly used as template for PCR. Olig2F and OligTSSR were used as primers to amplify the 2 kb fragment. Olig2.5 was added to see its inhibition to various DNAP. DNAP: DNA polymerase. LA: LA Taq DNAP. Q5: Q5 High-Fidelity DNAP. PS: PrimeSTAR HS DNAP. GXL: PrimeSTAR GXL DNAP. Phusion: Phusion High-Fidelity DNAP. Cobuddy: Cobuddy Super Fidelity DNAP. KOD: KOD DNAP.

    Journal: Scientific Reports

    Article Title: TT(N)mGCCTC inhibits archaeal family B DNA polymerases

    doi: 10.1038/s41598-018-20127-4

    Figure Lengend Snippet: Olig2.5 inhibited archaeal family B DNA polymerases. An E. coli DH5α colony harbouring the 2.6 kb upstream sequence of Olig2 in pGL3-Basic vector was cultured and 1 μl of the culture was directly used as template for PCR. Olig2F and OligTSSR were used as primers to amplify the 2 kb fragment. Olig2.5 was added to see its inhibition to various DNAP. DNAP: DNA polymerase. LA: LA Taq DNAP. Q5: Q5 High-Fidelity DNAP. PS: PrimeSTAR HS DNAP. GXL: PrimeSTAR GXL DNAP. Phusion: Phusion High-Fidelity DNAP. Cobuddy: Cobuddy Super Fidelity DNAP. KOD: KOD DNAP.

    Article Snippet: Phusion® High-Fidelity DNA Polymerase (Phusion) and Q5® High-Fidelity DNA Polymerase (Q5) were from New England Biolabs.

    Techniques: Sequencing, Plasmid Preparation, Cell Culture, Polymerase Chain Reaction, Inhibition

    Assay specificity for PIK 3 CA mutations. Comparison of mutant PIK 3 CA detection in human control genomic DNA ( gDNA ) and cell‐free plasma DNA following pre‐amplification or no pre‐amplification using Q5 High‐Fidelity DNA Polymerase (New England BioLabs) or TaqMan ® PreAmp Master Mix (Life Technologies). The level of blank (LoB) was 0.04% for the mutation E545K and for H1047R, 0.02% for E542K, and less than 0.01% for H1047L as shown by the graphs.

    Journal: Molecular Oncology

    Article Title: Correlation between circulating cell‐free PIK3CA tumor DNA levels and treatment response in patients with PIK3CA‐mutated metastatic breast cancer

    doi: 10.1002/1878-0261.12305

    Figure Lengend Snippet: Assay specificity for PIK 3 CA mutations. Comparison of mutant PIK 3 CA detection in human control genomic DNA ( gDNA ) and cell‐free plasma DNA following pre‐amplification or no pre‐amplification using Q5 High‐Fidelity DNA Polymerase (New England BioLabs) or TaqMan ® PreAmp Master Mix (Life Technologies). The level of blank (LoB) was 0.04% for the mutation E545K and for H1047R, 0.02% for E542K, and less than 0.01% for H1047L as shown by the graphs.

    Article Snippet: To increase the sensitivity of the analysis, the remaining 75 μL serum DNA was subjected to 12 cycles of PCR pre‐amplification with Q5 High‐Fidelity DNA Polymerase (New England BioLabs, Ipswich, MA, USA) using a multiplex PIK3CA primer mix.

    Techniques: Mutagenesis, Amplification

    Tomato fruit colour DSB repair assay. ( a ) Crossing yellow flesh e 3756 35S:Cas9 and bicolor cc383 u6-26:Ps#1-sgRNA gives F 1 plants with a pale Bicolour fruit phenotype. F 1 plants expressing both Cas9 and gRNA were selected. The gRNA was designed for DSB induction (black lightning) in both alleles between the yellow flesh e 3756 and bicolor cc383 mutations (*). In case of error-prone NHEJ repair (blue line) of bicolor cc383 , fruit colour is yellow. In cases of non-crossover or crossover, fruit colour is expected to be red or bicolour or yellow with red spots in case of late event. Note that whole red fruits were not obtained. Rather, fruits with red spots in a yellow or bicolour background were found and are shown together with additional products of HR-induced repair in Supplementary Fig. 1 . ( b ) Fruit phenotype distribution in F 1 plants and control: Bicolour fruits are shown as orange boxes; Yellow fruits as yellow; Fruits with red sectors (putative somatic HR) are shown as red-stripped boxes. Each column represents a fruit population derived from cross of independent transgenic lines of Cas9 and a given u6-26:Ps#1-sgRNA line. The number of fruits analysed is shown on the column in black. ( c ) Sequences of the NHEJ DSB repair footprints and their relative frequency are shown. The CRISPR-Cas target sequence from the PSY1 is shown on the top. The PSY1 start codon is shown in red and the PAM in blue. The top pie represents an average of illumina Hiseq reads from 22 different F 1 plants of the cross of yellow flesh e 3756 35S:Cas9 × bicolor cc383 u6-26:Ps#1-sgRNA. The low pie represents an average of ilummina Hiseq reads from two plants of control F 1 population ( yellow flesh e 3756 × bicolor cc383 F 1 plants with no CRISPR-Cas). ( d ) Inverse PCR scheme for identification of recombinant DNA fragments (details in Supplementary Fig. 4 ). (1) DNA from separate leaves was digested with ApaI(A) and HindIII(H) and then blunted. (2) Each sample was self-ligated, and (3) amplified by two different primer sets (green and yellow). Blue- Bicolor ; red- Yellow flesh ; Dashed blue line- Bicolor deletion, *- Yellow flesh mutation. ( e ) Ratio of parental (P) versus recombinant (R) types (obtained from panel C) in individual plants. Plants 1–15- F 1 plants of the cross of yellow flesh e 3756 35S:Cas9 × bicolor cc383 u6-26:Ps#1-sgRNA; Plant 16- synthetic crossover control; Plants17–18-Yellow flesh × Bicolor (Cas9-) F 1 plants.

    Journal: Nature Communications

    Article Title: Targeted recombination between homologous chromosomes for precise breeding in tomato

    doi: 10.1038/ncomms15605

    Figure Lengend Snippet: Tomato fruit colour DSB repair assay. ( a ) Crossing yellow flesh e 3756 35S:Cas9 and bicolor cc383 u6-26:Ps#1-sgRNA gives F 1 plants with a pale Bicolour fruit phenotype. F 1 plants expressing both Cas9 and gRNA were selected. The gRNA was designed for DSB induction (black lightning) in both alleles between the yellow flesh e 3756 and bicolor cc383 mutations (*). In case of error-prone NHEJ repair (blue line) of bicolor cc383 , fruit colour is yellow. In cases of non-crossover or crossover, fruit colour is expected to be red or bicolour or yellow with red spots in case of late event. Note that whole red fruits were not obtained. Rather, fruits with red spots in a yellow or bicolour background were found and are shown together with additional products of HR-induced repair in Supplementary Fig. 1 . ( b ) Fruit phenotype distribution in F 1 plants and control: Bicolour fruits are shown as orange boxes; Yellow fruits as yellow; Fruits with red sectors (putative somatic HR) are shown as red-stripped boxes. Each column represents a fruit population derived from cross of independent transgenic lines of Cas9 and a given u6-26:Ps#1-sgRNA line. The number of fruits analysed is shown on the column in black. ( c ) Sequences of the NHEJ DSB repair footprints and their relative frequency are shown. The CRISPR-Cas target sequence from the PSY1 is shown on the top. The PSY1 start codon is shown in red and the PAM in blue. The top pie represents an average of illumina Hiseq reads from 22 different F 1 plants of the cross of yellow flesh e 3756 35S:Cas9 × bicolor cc383 u6-26:Ps#1-sgRNA. The low pie represents an average of ilummina Hiseq reads from two plants of control F 1 population ( yellow flesh e 3756 × bicolor cc383 F 1 plants with no CRISPR-Cas). ( d ) Inverse PCR scheme for identification of recombinant DNA fragments (details in Supplementary Fig. 4 ). (1) DNA from separate leaves was digested with ApaI(A) and HindIII(H) and then blunted. (2) Each sample was self-ligated, and (3) amplified by two different primer sets (green and yellow). Blue- Bicolor ; red- Yellow flesh ; Dashed blue line- Bicolor deletion, *- Yellow flesh mutation. ( e ) Ratio of parental (P) versus recombinant (R) types (obtained from panel C) in individual plants. Plants 1–15- F 1 plants of the cross of yellow flesh e 3756 35S:Cas9 × bicolor cc383 u6-26:Ps#1-sgRNA; Plant 16- synthetic crossover control; Plants17–18-Yellow flesh × Bicolor (Cas9-) F 1 plants.

    Article Snippet: High-Fidelity DNA polymerase (New England BioLabs) and 18 PCR cycles (for specific primers of each experiment see primers list).

    Techniques: Expressing, Non-Homologous End Joining, Derivative Assay, Transgenic Assay, CRISPR, Sequencing, Inverse PCR, Recombinant, Amplification, Mutagenesis

    Optimization of EvaGreen- and Q5 DNA polymerase-based dNTP quantification. ( A-F ) Each critical reaction component and reaction temperature were varied and the outcome evaluated. ( G ) Assay conditions in graphs A to F . The standard curves were generated from end-point (1h) baseline-corrected fluorescence values.

    Journal: bioRxiv

    Article Title: A sensitive assay for dNTPs based on long synthetic oligonucleotides, EvaGreen dye, and inhibitor-resistant high-fidelity DNA polymerase

    doi: 10.1101/2019.12.17.879122

    Figure Lengend Snippet: Optimization of EvaGreen- and Q5 DNA polymerase-based dNTP quantification. ( A-F ) Each critical reaction component and reaction temperature were varied and the outcome evaluated. ( G ) Assay conditions in graphs A to F . The standard curves were generated from end-point (1h) baseline-corrected fluorescence values.

    Article Snippet: Based on the lowest background signal, and reported ( ) inhibitor resistance in DNA extraction-free PCR, we chose to continue the optimization with Q5 DNA polymerase.

    Techniques: Generated, Fluorescence

    Long synthetic DNA oligonucleotides, an inhibitor-resistant high-fidelity DNA polymerase and EvaGreen detection chemistry allow quantification of dNTPs from mouse liver extracts. ( A ) Representative quantification of dTTP using the published fluorometric probe-hydrolysis-based assay. For clarity, curves from three lowest standard samples (0.31 to 0.08 pmol) are not shown. ( B ) Representative quantification of dTTP using Q5 DNA polymerase, long oligonucleotide template and EvaGreen detection chemistry. ( C - D ) dTTP signal generated by the fluorometric methods from mouse liver extracts with and without 0.5 pmol dTTP spike-in calibrant. The final extract volume was diluted to 80 µl per 40 mg of initial tissue weight. ( E - F ) Standard curves generated from the end-point baseline-corrected fluorescence values. The lowest y-axis value shows the background signal. Gray lines present the 95% confidence interval of the non-linear curve fit.

    Journal: bioRxiv

    Article Title: A sensitive assay for dNTPs based on long synthetic oligonucleotides, EvaGreen dye, and inhibitor-resistant high-fidelity DNA polymerase

    doi: 10.1101/2019.12.17.879122

    Figure Lengend Snippet: Long synthetic DNA oligonucleotides, an inhibitor-resistant high-fidelity DNA polymerase and EvaGreen detection chemistry allow quantification of dNTPs from mouse liver extracts. ( A ) Representative quantification of dTTP using the published fluorometric probe-hydrolysis-based assay. For clarity, curves from three lowest standard samples (0.31 to 0.08 pmol) are not shown. ( B ) Representative quantification of dTTP using Q5 DNA polymerase, long oligonucleotide template and EvaGreen detection chemistry. ( C - D ) dTTP signal generated by the fluorometric methods from mouse liver extracts with and without 0.5 pmol dTTP spike-in calibrant. The final extract volume was diluted to 80 µl per 40 mg of initial tissue weight. ( E - F ) Standard curves generated from the end-point baseline-corrected fluorescence values. The lowest y-axis value shows the background signal. Gray lines present the 95% confidence interval of the non-linear curve fit.

    Article Snippet: Based on the lowest background signal, and reported ( ) inhibitor resistance in DNA extraction-free PCR, we chose to continue the optimization with Q5 DNA polymerase.

    Techniques: Hydrolysis Assay, Generated, Fluorescence

    Deoxynucleoside diphosphate utilization by DNA polymerases. ( A ) A standard PCR with a monophosphate substrate is inhibited, whereas the presence of all four di- or triphosphate nucleotides supports DNA amplification. Thermophilic polymerases Taq, Vent (exo−), and Pfu , as well as B , Deep Vent, and Q5 DNA polymerases, also utilize the diphosphorylated substrates. ( C and D ) Primer-extension reactions on short templates sampled at indicated time points. All reactions were performed with 100 µM dNDPs at 60 °C, except the Bsu reactions, which were performed at 37 °C. Extended sequences are shown alongside of gels and were distinct in C and D . Primer band is indicated with “–P.” Pausing appears mainly before incorporation of dA and dC and is variable among the polymerases. ( E ).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: DNA synthesis from diphosphate substrates by DNA polymerases

    doi: 10.1073/pnas.1712193115

    Figure Lengend Snippet: Deoxynucleoside diphosphate utilization by DNA polymerases. ( A ) A standard PCR with a monophosphate substrate is inhibited, whereas the presence of all four di- or triphosphate nucleotides supports DNA amplification. Thermophilic polymerases Taq, Vent (exo−), and Pfu , as well as B , Deep Vent, and Q5 DNA polymerases, also utilize the diphosphorylated substrates. ( C and D ) Primer-extension reactions on short templates sampled at indicated time points. All reactions were performed with 100 µM dNDPs at 60 °C, except the Bsu reactions, which were performed at 37 °C. Extended sequences are shown alongside of gels and were distinct in C and D . Primer band is indicated with “–P.” Pausing appears mainly before incorporation of dA and dC and is variable among the polymerases. ( E ).

    Article Snippet: Taq DNA polymerase, Phusion High-Fidelity DNA polymerase, DeepVent DNA polymerase, Vent DNA polymerase, Q5 DNA High-Fidelity polymerase, Bst polymerase, Bsu DNA polymerase (large fragment), and ThermoPol Reaction Buffer [1×: 20 mM Tris⋅HCl, 10 mM (NH4 )2 SO4 , 10 mM KCl, 2 mM MgSO4 , 0.1% Triton–X–100, pH 8.8 at 25 °C] were purchased from New England Biolabs.

    Techniques: Polymerase Chain Reaction, Amplification