q5 polymerase New England Biolabs Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs q5 high fidelity dna polymerase new england biolabs
    PCR based verification of genome edit A. A schematic representation showing the locations of PCR primers used in panel B; B. Agarose gel stained with ethidium bromide of amplified PCR products using primers specific to the point mutation (H743A, for which we changed CAC→GCT) and the B. subtilis chromosome (left panel), and using primers specific to the editing plasmid (right panel). Of twelve isolates screened all were found to have the predicted point mutation, and two of twelve retained resistance to spectinomycin (asterisks). The slow migrating band in the PY79 lane corresponds to nonspecific PCR amplification in the wild type background. The editing plasmid was detectable in spectinomycin resistant isolates, but not in the remaining ten. C. Agarose gel stained with ethidium bromide of PCR reactions using primers specific for deletion of four B. subtilis genes. Primers were designed outside of the editing template, and are therefore specific to chromosomal DNA. For all four deletions, all isolates were found to be spectinomycin sensitive, and PCR genotyping of three isolates for each deletion found that all isolates tested had the intended deletion. Note that for the control reaction for ponA , the PCR product is faint because the amplicon is approximately 5 kb and the extension time of the PCR was 3 min, which is near the limit for complete extension for <t>Q5</t> DNA polymerase; the other three control amplicons are no larger than 3.4 kb.
    Q5 High Fidelity Dna Polymerase New England Biolabs, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 910 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 high fidelity dna polymerase new england biolabs/product/New England Biolabs
    Average 99 stars, based on 910 article reviews
    Price from $9.99 to $1999.99
    q5 high fidelity dna polymerase new england biolabs - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    80
    New England Biolabs q5 hot start high fidelty dna polymerase new england biolabs cat
    PCR based verification of genome edit A. A schematic representation showing the locations of PCR primers used in panel B; B. Agarose gel stained with ethidium bromide of amplified PCR products using primers specific to the point mutation (H743A, for which we changed CAC→GCT) and the B. subtilis chromosome (left panel), and using primers specific to the editing plasmid (right panel). Of twelve isolates screened all were found to have the predicted point mutation, and two of twelve retained resistance to spectinomycin (asterisks). The slow migrating band in the PY79 lane corresponds to nonspecific PCR amplification in the wild type background. The editing plasmid was detectable in spectinomycin resistant isolates, but not in the remaining ten. C. Agarose gel stained with ethidium bromide of PCR reactions using primers specific for deletion of four B. subtilis genes. Primers were designed outside of the editing template, and are therefore specific to chromosomal DNA. For all four deletions, all isolates were found to be spectinomycin sensitive, and PCR genotyping of three isolates for each deletion found that all isolates tested had the intended deletion. Note that for the control reaction for ponA , the PCR product is faint because the amplicon is approximately 5 kb and the extension time of the PCR was 3 min, which is near the limit for complete extension for <t>Q5</t> DNA polymerase; the other three control amplicons are no larger than 3.4 kb.
    Q5 Hot Start High Fidelty Dna Polymerase New England Biolabs Cat, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 hot start high fidelty dna polymerase new england biolabs cat/product/New England Biolabs
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    q5 hot start high fidelty dna polymerase new england biolabs cat - by Bioz Stars, 2020-04
    80/100 stars
      Buy from Supplier

    99
    New England Biolabs new england biolabs q5 high fidelity
    PCR based verification of genome edit A. A schematic representation showing the locations of PCR primers used in panel B; B. Agarose gel stained with ethidium bromide of amplified PCR products using primers specific to the point mutation (H743A, for which we changed CAC→GCT) and the B. subtilis chromosome (left panel), and using primers specific to the editing plasmid (right panel). Of twelve isolates screened all were found to have the predicted point mutation, and two of twelve retained resistance to spectinomycin (asterisks). The slow migrating band in the PY79 lane corresponds to nonspecific PCR amplification in the wild type background. The editing plasmid was detectable in spectinomycin resistant isolates, but not in the remaining ten. C. Agarose gel stained with ethidium bromide of PCR reactions using primers specific for deletion of four B. subtilis genes. Primers were designed outside of the editing template, and are therefore specific to chromosomal DNA. For all four deletions, all isolates were found to be spectinomycin sensitive, and PCR genotyping of three isolates for each deletion found that all isolates tested had the intended deletion. Note that for the control reaction for ponA , the PCR product is faint because the amplicon is approximately 5 kb and the extension time of the PCR was 3 min, which is near the limit for complete extension for <t>Q5</t> DNA polymerase; the other three control amplicons are no larger than 3.4 kb.
    New England Biolabs Q5 High Fidelity, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/new england biolabs q5 high fidelity/product/New England Biolabs
    Average 99 stars, based on 109 article reviews
    Price from $9.99 to $1999.99
    new england biolabs q5 high fidelity - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    New England Biolabs q5 reaction buffer
    PCR based verification of genome edit A. A schematic representation showing the locations of PCR primers used in panel B; B. Agarose gel stained with ethidium bromide of amplified PCR products using primers specific to the point mutation (H743A, for which we changed CAC→GCT) and the B. subtilis chromosome (left panel), and using primers specific to the editing plasmid (right panel). Of twelve isolates screened all were found to have the predicted point mutation, and two of twelve retained resistance to spectinomycin (asterisks). The slow migrating band in the PY79 lane corresponds to nonspecific PCR amplification in the wild type background. The editing plasmid was detectable in spectinomycin resistant isolates, but not in the remaining ten. C. Agarose gel stained with ethidium bromide of PCR reactions using primers specific for deletion of four B. subtilis genes. Primers were designed outside of the editing template, and are therefore specific to chromosomal DNA. For all four deletions, all isolates were found to be spectinomycin sensitive, and PCR genotyping of three isolates for each deletion found that all isolates tested had the intended deletion. Note that for the control reaction for ponA , the PCR product is faint because the amplicon is approximately 5 kb and the extension time of the PCR was 3 min, which is near the limit for complete extension for <t>Q5</t> DNA polymerase; the other three control amplicons are no larger than 3.4 kb.
    Q5 Reaction Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 595 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 reaction buffer/product/New England Biolabs
    Average 99 stars, based on 595 article reviews
    Price from $9.99 to $1999.99
    q5 reaction buffer - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    New England Biolabs q5 hot start polymerase
    Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using <t>Q5</t> Hot Start High-Fidelity <t>DNA</t> polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)
    Q5 Hot Start Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 733 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 hot start polymerase/product/New England Biolabs
    Average 99 stars, based on 733 article reviews
    Price from $9.99 to $1999.99
    q5 hot start polymerase - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    90
    New England Biolabs nebnext q5
    Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using <t>Q5</t> Hot Start High-Fidelity <t>DNA</t> polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)
    Nebnext Q5, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nebnext q5/product/New England Biolabs
    Average 90 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    nebnext q5 - by Bioz Stars, 2020-04
    90/100 stars
      Buy from Supplier

    99
    New England Biolabs ultratm ii q5
    Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using <t>Q5</t> Hot Start High-Fidelity <t>DNA</t> polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)
    Ultratm Ii Q5, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ultratm ii q5/product/New England Biolabs
    Average 99 stars, based on 19 article reviews
    Price from $9.99 to $1999.99
    ultratm ii q5 - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    94
    Promega q5 polymerases
    Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using <t>Q5</t> Hot Start High-Fidelity <t>DNA</t> polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)
    Q5 Polymerases, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 polymerases/product/Promega
    Average 94 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    q5 polymerases - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    99
    New England Biolabs q5 site directed mutagenesis kit
    Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using <t>Q5</t> Hot Start High-Fidelity <t>DNA</t> polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)
    Q5 Site Directed Mutagenesis Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 10153 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 site directed mutagenesis kit/product/New England Biolabs
    Average 99 stars, based on 10153 article reviews
    Price from $9.99 to $1999.99
    q5 site directed mutagenesis kit - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    New England Biolabs q5 hot start high fidelity
    Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using <t>Q5</t> Hot Start High-Fidelity <t>DNA</t> polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)
    Q5 Hot Start High Fidelity, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 534 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 hot start high fidelity/product/New England Biolabs
    Average 99 stars, based on 534 article reviews
    Price from $9.99 to $1999.99
    q5 hot start high fidelity - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    80
    New England Biolabs l neb q5 polymerase reaction
    Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using <t>Q5</t> Hot Start High-Fidelity <t>DNA</t> polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)
    L Neb Q5 Polymerase Reaction, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/l neb q5 polymerase reaction/product/New England Biolabs
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    l neb q5 polymerase reaction - by Bioz Stars, 2020-04
    80/100 stars
      Buy from Supplier

    99
    New England Biolabs q5 high fidelity pcr kit
    Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using <t>Q5</t> Hot Start High-Fidelity <t>DNA</t> polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)
    Q5 High Fidelity Pcr Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 300 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 high fidelity pcr kit/product/New England Biolabs
    Average 99 stars, based on 300 article reviews
    Price from $9.99 to $1999.99
    q5 high fidelity pcr kit - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    Image Search Results


    PCR based verification of genome edit A. A schematic representation showing the locations of PCR primers used in panel B; B. Agarose gel stained with ethidium bromide of amplified PCR products using primers specific to the point mutation (H743A, for which we changed CAC→GCT) and the B. subtilis chromosome (left panel), and using primers specific to the editing plasmid (right panel). Of twelve isolates screened all were found to have the predicted point mutation, and two of twelve retained resistance to spectinomycin (asterisks). The slow migrating band in the PY79 lane corresponds to nonspecific PCR amplification in the wild type background. The editing plasmid was detectable in spectinomycin resistant isolates, but not in the remaining ten. C. Agarose gel stained with ethidium bromide of PCR reactions using primers specific for deletion of four B. subtilis genes. Primers were designed outside of the editing template, and are therefore specific to chromosomal DNA. For all four deletions, all isolates were found to be spectinomycin sensitive, and PCR genotyping of three isolates for each deletion found that all isolates tested had the intended deletion. Note that for the control reaction for ponA , the PCR product is faint because the amplicon is approximately 5 kb and the extension time of the PCR was 3 min, which is near the limit for complete extension for Q5 DNA polymerase; the other three control amplicons are no larger than 3.4 kb.

    Journal: Bio-protocol

    Article Title: CRISPR/Cas9 Editing of the Bacillus subtilis Genome

    doi: 10.21769/BioProtoc.2272

    Figure Lengend Snippet: PCR based verification of genome edit A. A schematic representation showing the locations of PCR primers used in panel B; B. Agarose gel stained with ethidium bromide of amplified PCR products using primers specific to the point mutation (H743A, for which we changed CAC→GCT) and the B. subtilis chromosome (left panel), and using primers specific to the editing plasmid (right panel). Of twelve isolates screened all were found to have the predicted point mutation, and two of twelve retained resistance to spectinomycin (asterisks). The slow migrating band in the PY79 lane corresponds to nonspecific PCR amplification in the wild type background. The editing plasmid was detectable in spectinomycin resistant isolates, but not in the remaining ten. C. Agarose gel stained with ethidium bromide of PCR reactions using primers specific for deletion of four B. subtilis genes. Primers were designed outside of the editing template, and are therefore specific to chromosomal DNA. For all four deletions, all isolates were found to be spectinomycin sensitive, and PCR genotyping of three isolates for each deletion found that all isolates tested had the intended deletion. Note that for the control reaction for ponA , the PCR product is faint because the amplicon is approximately 5 kb and the extension time of the PCR was 3 min, which is near the limit for complete extension for Q5 DNA polymerase; the other three control amplicons are no larger than 3.4 kb.

    Article Snippet: The PCR program used to linearize pPB41 is: PCR amplify CRISPR/Cas9 using plasmid generated in step B1 ( ) Amplify plasmid generated in step B1 (pPB43 in the example) via PCR using Q5 DNA polymerase (NEB) and primers oPEB232 and oPEB234.

    Techniques: Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Amplification, Mutagenesis, Plasmid Preparation

    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

    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

    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

    Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using Q5 Hot Start High-Fidelity DNA polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)

    Journal: Proceedings of the Royal Society B: Biological Sciences

    Article Title: Disentangling sources of variation in SSU rDNA sequences from single cell analyses of ciliates: impact of copy number variation and experimental error

    doi: 10.1098/rspb.2017.0425

    Figure Lengend Snippet: Distribution of polymorphic sites in the SSU rDNA of Halteria grandinella , Blepharisma americanum and Strombidium stylifer amplified using Q5 Hot Start High-Fidelity DNA polymerase. Polymorphic sites are indicated by short vertical lines. The squares, circles and triangles represent the three individuals of each species. The two-way arrows indicate the hypervariable regions of SSU rDNA and the lengths of sequences are to scale. Hal, Halteria grandinella ; Ble, Blepharisma americanum ; Str, Strombidium stylifer . (Online version in colour.)

    Article Snippet: Afterwards, PfuTurbo DNA polymerase (Cat. #600250, Agilent Technologies, USA), Q5 Hot Start High-Fidelity DNA Polymerase (Cat. #M0493 L, New England Biolabs, USA), ExTaq DNA polymerase (Cat. #RR001A & #RR003A, TaKaRa, Japan) and Taq DNA Polymerase (Cat. #EP0402, Thermo Fisher Scientific, USA) were used to amplify the SSU rDNA in the plasmid with universal primers [ ].

    Techniques: Amplification

    Performance of four thermostable DNA polymerases for the synthesis of gene B using PCA-DTF. Lane 1: negative control reaction performed at the same conditions, without addition of primers; lane 2: KOD Hot Start DNA polymerase; lane 3: Q5 Hot Start High Fidelity DNA polymerase; lane 4: Pfu Turbo DNA polymerase and lane 5: Taq DNA polymerase. M2: NZYladder I

    Journal: BMC Biotechnology

    Article Title: Development of a gene synthesis platform for the efficient large scale production of small genes encoding animal toxins

    doi: 10.1186/s12896-016-0316-3

    Figure Lengend Snippet: Performance of four thermostable DNA polymerases for the synthesis of gene B using PCA-DTF. Lane 1: negative control reaction performed at the same conditions, without addition of primers; lane 2: KOD Hot Start DNA polymerase; lane 3: Q5 Hot Start High Fidelity DNA polymerase; lane 4: Pfu Turbo DNA polymerase and lane 5: Taq DNA polymerase. M2: NZYladder I

    Article Snippet: Finally, extension was performed at 70 °C for 3 s for KOD Hot Start DNA polymerase, 72 °C for 30 s for Pfu Turbo and Taq, and 72 °C for 15 s for Q5® Hot Star High Fidelity DNA polymerase.

    Techniques: Negative Control