standard taq reaction buffer  (New England Biolabs)


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    Name:
    Standard Taq Reaction Buffer Pack
    Description:
    Standard Taq Reaction Buffer Pack 6 0 ml
    Catalog Number:
    b9014s
    Price:
    22
    Size:
    6 0 ml
    Category:
    Buffers
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    Structured Review

    New England Biolabs standard taq reaction buffer
    Standard Taq Reaction Buffer Pack
    Standard Taq Reaction Buffer Pack 6 0 ml
    https://www.bioz.com/result/standard taq reaction buffer/product/New England Biolabs
    Average 99 stars, based on 23 article reviews
    Price from $9.99 to $1999.99
    standard taq reaction buffer - by Bioz Stars, 2020-05
    99/100 stars

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    Polymerase Chain Reaction:

    Article Title: Independently founded populations of Sclerotinia sclerotiorum from a tropical and a temperate region have similar genetic structure
    Article Snippet: .. PCR reactions were performed in a final volume of 20 μl with 2 μl of DNA, 0.5 μl of each primer at 10 mM, 0.5 μl of dNTPs at 10 mM, 2 μl of the 10x Standard Taq Reaction Buffer that includes 1.5 mM MgCl2 , 10mM Tris-HCl and 50 mM KCl (New England Biolabs Inc., Ipswich, MA, USA) and 0.1 μl of Taq DNA polymerase (New England Biolabs, Inc.). .. PCR reactions were conducted in a C1000 Thermal Cycler (Bio-Rad, Hercules, CA, USA).

    Article Title: Breaking the speed limit with multimode fast scanning of DNA by Endonuclease V
    Article Snippet: .. 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.

    Article Title: High intralocus variability and interlocus recombination promote immunological diversity in a minimal major histocompatibility system
    Article Snippet: .. PCR reactions for ACTB were performed in 20 μl volumes containing 0.5U Taq (New England Biolabs), 1× NEB reaction buffer, 0.75 μM MgCl2 , 0.2 mM dNTPs, 0.2 μM primers, and 1.0 μl DNA, with the following PCR cycling conditions: 95°C for 1:30, then 30 cycles of 95°C for 0:30, 53°C for 0:30, 68°C for 1:00, followed by 68°C for 5:00. .. MH amplifications were performed in 25 μl volumes containing 1U Taq (NEB), 1× NEB reaction buffer, 1.0 μM MgCl2 , 0.4 mM dNTPs, 0.2 μM primers and 2.5 μl DNA, with the following cycling parameters: 92°C for 0:10, then 40 cycles of 92°C for 0:10, 55°C for 0:30, 68°C for 2:00 (for MH IIα) or 92°C for 5:00, then 40 cycles of 92°C for 0:30, 62°C for 0:30, 68°C for 4:00, followed by 68°C for 15:00 (for MH IIβ).

    Article Title: Genetic Dissection of Tropodithietic Acid Biosynthesis by Marine Roseobacters ▿Genetic Dissection of Tropodithietic Acid Biosynthesis by Marine Roseobacters ▿ ‡
    Article Snippet: .. The standard PCR amplification conditions were 100 μM concentrations of each deoxynucleoside triphosphate, 0.2 μM concentrations of each primer, and 1 U of Taq DNA polymerase (New England Biolabs) in 1× reaction buffer (New England Biolabs) with an initial denaturing step at 94°C for 3 min, followed by 30 cycles of 94°C for 1 min each, annealing at 55°C for 30 s, and an elongation at 72°C for 1 min. To detect the tdaA-E locus, PCR amplification was conducted with a forward primer complementary to tdaA (5′-CGCTTTCCGGAACTGGAGAT-3′) and a reverse primer complementary to tdaE (5′-GGCTGCCGTATAGTTTCAGCA-3′) using the Expand Long Template PCR system (Roche Applied Science, Indianapolis, IN), and the PCR program conditions and cycle parameters were as described by the supplier. .. DNA-DNA hybridization by Southern slot blot ( ) was used to detect the presence of tda genes in other roseobacters.

    Article Title: Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression
    Article Snippet: .. The amplified fragment was purified by Qiaquick PCR purification (Qiagen) and incubated with five units of standard Taq polymerase in standard Taq buffer with 2.5 mM dATP (NEB) for 15 min at 72 °C to add A overhangs. .. This was then TA cloned using a TOPO® TA Cloning® Kit (Thermo Fisher Scientific).

    Article Title: DNA barcoding and LC-MS metabolite profiling of the lichen-forming genus Melanelia: Specimen identification and discrimination focusing on Icelandic taxa
    Article Snippet: .. Each amplification reaction (25 μL) contained 1×standard Taq reaction buffer for nrITS, 200 μM dNTPs, 0.2 μM forward and reverse primer, 1.25 units of Taq DNA polymerase (New England Biolabs), 1 or 3 μL DNA template, and PCR-grade water. .. PCR cycling conditions were as follows: initial denaturation at 94°C for 3 min, 31 cycles of 94°C for 40s, 54°C for 40s and 68°C for 1min, final extension at 68°C for 7min and cooling down to 4°C.

    Isolation:

    Article Title: Early Control of Mycobacterium tuberculosis Infection Requires il12rb1 Expression by rag1-Dependent Lineages
    Article Snippet: .. To screen for the presence of a wild-type il12rb1 allele, genomic DNA (gDNA) from tail snips was isolated using the Promega Wizard SV Genomic DNA method (Promega, Madison, WI) and amplified with primers NF1 (5′-CAGAGATCCTCCTGCCTCTG-3′) and NF2 (5′-TATGGTTCGGAGGGACAAAG-3′) in a 50-μl reaction mixture comprising 2 μl of gDNA, 1.5× NEB Taq buffer (from a 10× solution), 2 mM MgCl2 , 0.2 μM primer NF1, 0.2 μM primer NF2, 0.2 mM each deoxynucleoside triphosphate (dNTP), 5% dimethyl sulfoxide (DMSO), and 1U of Taq polymerase (New England BioLabs, Ipswich, MA). .. To screen for the knockout (KO) il12rb1 allele, gDNA was amplified with primers NF1 (sequence above) and NF3 (5′-TGGATGTGGAATGTGTGCGAG-3′) in a 50-μl reaction mixture comprising 2 μl of gDNA, 1.0× NEB Taq buffer (from a 10× solution), 2 mM MgCl2 , 0.2 μM primer NF1, 0.2 μM primer NF3, 0.2 mM each dNTP, 5% DMSO, and 1U of Taq polymerase.

    Incubation:

    Article Title: Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression
    Article Snippet: .. The amplified fragment was purified by Qiaquick PCR purification (Qiagen) and incubated with five units of standard Taq polymerase in standard Taq buffer with 2.5 mM dATP (NEB) for 15 min at 72 °C to add A overhangs. .. This was then TA cloned using a TOPO® TA Cloning® Kit (Thermo Fisher Scientific).

    Amplification:

    Article Title: Early Control of Mycobacterium tuberculosis Infection Requires il12rb1 Expression by rag1-Dependent Lineages
    Article Snippet: .. To screen for the presence of a wild-type il12rb1 allele, genomic DNA (gDNA) from tail snips was isolated using the Promega Wizard SV Genomic DNA method (Promega, Madison, WI) and amplified with primers NF1 (5′-CAGAGATCCTCCTGCCTCTG-3′) and NF2 (5′-TATGGTTCGGAGGGACAAAG-3′) in a 50-μl reaction mixture comprising 2 μl of gDNA, 1.5× NEB Taq buffer (from a 10× solution), 2 mM MgCl2 , 0.2 μM primer NF1, 0.2 μM primer NF2, 0.2 mM each deoxynucleoside triphosphate (dNTP), 5% dimethyl sulfoxide (DMSO), and 1U of Taq polymerase (New England BioLabs, Ipswich, MA). .. To screen for the knockout (KO) il12rb1 allele, gDNA was amplified with primers NF1 (sequence above) and NF3 (5′-TGGATGTGGAATGTGTGCGAG-3′) in a 50-μl reaction mixture comprising 2 μl of gDNA, 1.0× NEB Taq buffer (from a 10× solution), 2 mM MgCl2 , 0.2 μM primer NF1, 0.2 μM primer NF3, 0.2 mM each dNTP, 5% DMSO, and 1U of Taq polymerase.

    Article Title: Genetic Dissection of Tropodithietic Acid Biosynthesis by Marine Roseobacters ▿Genetic Dissection of Tropodithietic Acid Biosynthesis by Marine Roseobacters ▿ ‡
    Article Snippet: .. The standard PCR amplification conditions were 100 μM concentrations of each deoxynucleoside triphosphate, 0.2 μM concentrations of each primer, and 1 U of Taq DNA polymerase (New England Biolabs) in 1× reaction buffer (New England Biolabs) with an initial denaturing step at 94°C for 3 min, followed by 30 cycles of 94°C for 1 min each, annealing at 55°C for 30 s, and an elongation at 72°C for 1 min. To detect the tdaA-E locus, PCR amplification was conducted with a forward primer complementary to tdaA (5′-CGCTTTCCGGAACTGGAGAT-3′) and a reverse primer complementary to tdaE (5′-GGCTGCCGTATAGTTTCAGCA-3′) using the Expand Long Template PCR system (Roche Applied Science, Indianapolis, IN), and the PCR program conditions and cycle parameters were as described by the supplier. .. DNA-DNA hybridization by Southern slot blot ( ) was used to detect the presence of tda genes in other roseobacters.

    Article Title: Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression
    Article Snippet: .. The amplified fragment was purified by Qiaquick PCR purification (Qiagen) and incubated with five units of standard Taq polymerase in standard Taq buffer with 2.5 mM dATP (NEB) for 15 min at 72 °C to add A overhangs. .. This was then TA cloned using a TOPO® TA Cloning® Kit (Thermo Fisher Scientific).

    Article Title: DNA barcoding and LC-MS metabolite profiling of the lichen-forming genus Melanelia: Specimen identification and discrimination focusing on Icelandic taxa
    Article Snippet: .. Each amplification reaction (25 μL) contained 1×standard Taq reaction buffer for nrITS, 200 μM dNTPs, 0.2 μM forward and reverse primer, 1.25 units of Taq DNA polymerase (New England Biolabs), 1 or 3 μL DNA template, and PCR-grade water. .. PCR cycling conditions were as follows: initial denaturation at 94°C for 3 min, 31 cycles of 94°C for 40s, 54°C for 40s and 68°C for 1min, final extension at 68°C for 7min and cooling down to 4°C.

    Purification:

    Article Title: Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression
    Article Snippet: .. The amplified fragment was purified by Qiaquick PCR purification (Qiagen) and incubated with five units of standard Taq polymerase in standard Taq buffer with 2.5 mM dATP (NEB) for 15 min at 72 °C to add A overhangs. .. This was then TA cloned using a TOPO® TA Cloning® Kit (Thermo Fisher Scientific).

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    New England Biolabs taq buffer
    PCR products of UBP4′: M-marker (bp) GeneRuler <t>DNA</t> Ladder Mix (Fermentas Life Sciences), lanes 1–8 UBP4′ gene (276 bp UBP4′ length and 26 bp for restriction enzymes sequence = 302 bp) PCR product: lanes 1, 2: PCR reaction with Biotools DNA polymerase and 23 cycles, lanes 3, 4: PCR reaction with Biotools DNA polymerase and 29 cycles, lanes 5, 6: PCR reaction with Biotools DNA polymerase and 29 cycles, and lanes 7, 8: PCR reaction with <t>Taq</t> DNA polymerase and 29 cycles.
    Taq Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 546 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taq buffer/product/New England Biolabs
    Average 99 stars, based on 546 article reviews
    Price from $9.99 to $1999.99
    taq buffer - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

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    PCR products of UBP4′: M-marker (bp) GeneRuler DNA Ladder Mix (Fermentas Life Sciences), lanes 1–8 UBP4′ gene (276 bp UBP4′ length and 26 bp for restriction enzymes sequence = 302 bp) PCR product: lanes 1, 2: PCR reaction with Biotools DNA polymerase and 23 cycles, lanes 3, 4: PCR reaction with Biotools DNA polymerase and 29 cycles, lanes 5, 6: PCR reaction with Biotools DNA polymerase and 29 cycles, and lanes 7, 8: PCR reaction with Taq DNA polymerase and 29 cycles.

    Journal: BioMed Research International

    Article Title: DNASynth: A Computer Program for Assembly of Artificial Gene Parts in Decreasing Temperature

    doi: 10.1155/2015/413262

    Figure Lengend Snippet: PCR products of UBP4′: M-marker (bp) GeneRuler DNA Ladder Mix (Fermentas Life Sciences), lanes 1–8 UBP4′ gene (276 bp UBP4′ length and 26 bp for restriction enzymes sequence = 302 bp) PCR product: lanes 1, 2: PCR reaction with Biotools DNA polymerase and 23 cycles, lanes 3, 4: PCR reaction with Biotools DNA polymerase and 29 cycles, lanes 5, 6: PCR reaction with Biotools DNA polymerase and 29 cycles, and lanes 7, 8: PCR reaction with Taq DNA polymerase and 29 cycles.

    Article Snippet: S.A. or Taq DNA polymerase with standard Taq buffer, New England Biolabs, Inc.), and 1 μ L as a template for 23 and 29 cycles using Eppendorf 5330 thermocycler.

    Techniques: Polymerase Chain Reaction, Marker, Sequencing

    CE analysis of processing synthetic DNA by soluble enzyme mix PKT and immobilized enzymes. 5′ FAM-labeled blunt-end substrates, 51-AT possessing multiple 3′ terminal A-T base pairs, and 51-GC possessing multiple 3′ terminal G-C base pairs, were incubated with PKT for end repair at 20 °C for 30 min followed by 65 °C for 30 min (PKT mix). The substrates were also treated with immobilized T4 DNA pol and PNK at 20 °C for 30 min, followed by separation of the enzymes on beads and the reaction medium (supernatant). The reaction medium was subsequently treated with immobilized Taq DNA pol for 3′ A-tailing at 37 °C for 30 min (IM PKT mix). The CE data show that incubation with PKT resulted in extensive degradation of 51-AT and little degradation of 51-GC. Treatment of 51-AT or 51-GC with the immobilized enzymes resulted in mostly 3′ A-tailing product, without detectable degradation of the 5′ FAM-labeled oligos. NC, negative control reaction performed in the absence of enzyme.

    Journal: Scientific Reports

    Article Title: Solid-phase enzyme catalysis of DNA end repair and 3′ A-tailing reduces GC-bias in next-generation sequencing of human genomic DNA

    doi: 10.1038/s41598-018-34079-2

    Figure Lengend Snippet: CE analysis of processing synthetic DNA by soluble enzyme mix PKT and immobilized enzymes. 5′ FAM-labeled blunt-end substrates, 51-AT possessing multiple 3′ terminal A-T base pairs, and 51-GC possessing multiple 3′ terminal G-C base pairs, were incubated with PKT for end repair at 20 °C for 30 min followed by 65 °C for 30 min (PKT mix). The substrates were also treated with immobilized T4 DNA pol and PNK at 20 °C for 30 min, followed by separation of the enzymes on beads and the reaction medium (supernatant). The reaction medium was subsequently treated with immobilized Taq DNA pol for 3′ A-tailing at 37 °C for 30 min (IM PKT mix). The CE data show that incubation with PKT resulted in extensive degradation of 51-AT and little degradation of 51-GC. Treatment of 51-AT or 51-GC with the immobilized enzymes resulted in mostly 3′ A-tailing product, without detectable degradation of the 5′ FAM-labeled oligos. NC, negative control reaction performed in the absence of enzyme.

    Article Snippet: Enzyme mix PKT was comprised of approximately 1,200 units/ml T4 DNA polymerase, 2,000 units/ml T4 PNK and 2,000 units/ml Taq DNA polymerase (NEB) while PK contained T4 DNA polymerase and T4 PNK only.

    Techniques: Labeling, Incubation, Negative Control

    Effect of end repair and 3′ A-tailing at high temperature on GC-bias in Illumina reads from PCR-free human DNA libraries. ( a ) Comparison of GC-bias curves in duplicate libraries prepared by immobilized enzymes with 3′ A-tailing performed at 37 °C (IM 37 °C -1 and IM 37 °C -2, in blue) or 65 °C (IM 65 °C -1 and IM 65 °C -2, in green) revealed a dramatic effect of 3′ A-tailing at high temperature on sequence coverage of the AT-rich regions from human DNA libraries. ( b ) GC-bias curves were generated from two sets of duplicate libraries produced using the soluble enzyme mixture PKT with (PKT purify-1 and PKT purify-2) or without (PKT-1 and PKT-2) a purification step between end repair and high temperature incubation (with Taq DNA pol added to the samples following purification). Although some bias against AT-rich regions can be attributed to the end repair step, the elevated temperature contributes to the majority of the dropouts in the AT-rich regions. ( c ) Shown are the GC-bias curves from 4 sets of duplicate libraries produced by the method of soluble enzymes. Two sets of the duplicate libraries were purified after end repair with PK mixture and then treated at 37 °C with Klenow Fragment (3′-5′ exo − ) (red, Klenow 37 °C-1 and Klenow 37 °C-2) or Taq DNA pol (blue, Taq 37 °C-1 and Taq 37 °C-2). The other two duplicate sets were prepared using the high temperature treatment protocol either with (green, Taq 65 °C-1 and Taq 65 °C-2) or without (orange, PKT-1 and PKT-2) a purification step between end repair with PKT and treatment with Taq DNA pol at 65 °C for 30 min. ( d ) Comparison of library yield of the three sets described above with or without (PKT on the left) a purification step between end repair and 3′ A-tailing indicates that purification caused substantial loss of library DNA.

    Journal: Scientific Reports

    Article Title: Solid-phase enzyme catalysis of DNA end repair and 3′ A-tailing reduces GC-bias in next-generation sequencing of human genomic DNA

    doi: 10.1038/s41598-018-34079-2

    Figure Lengend Snippet: Effect of end repair and 3′ A-tailing at high temperature on GC-bias in Illumina reads from PCR-free human DNA libraries. ( a ) Comparison of GC-bias curves in duplicate libraries prepared by immobilized enzymes with 3′ A-tailing performed at 37 °C (IM 37 °C -1 and IM 37 °C -2, in blue) or 65 °C (IM 65 °C -1 and IM 65 °C -2, in green) revealed a dramatic effect of 3′ A-tailing at high temperature on sequence coverage of the AT-rich regions from human DNA libraries. ( b ) GC-bias curves were generated from two sets of duplicate libraries produced using the soluble enzyme mixture PKT with (PKT purify-1 and PKT purify-2) or without (PKT-1 and PKT-2) a purification step between end repair and high temperature incubation (with Taq DNA pol added to the samples following purification). Although some bias against AT-rich regions can be attributed to the end repair step, the elevated temperature contributes to the majority of the dropouts in the AT-rich regions. ( c ) Shown are the GC-bias curves from 4 sets of duplicate libraries produced by the method of soluble enzymes. Two sets of the duplicate libraries were purified after end repair with PK mixture and then treated at 37 °C with Klenow Fragment (3′-5′ exo − ) (red, Klenow 37 °C-1 and Klenow 37 °C-2) or Taq DNA pol (blue, Taq 37 °C-1 and Taq 37 °C-2). The other two duplicate sets were prepared using the high temperature treatment protocol either with (green, Taq 65 °C-1 and Taq 65 °C-2) or without (orange, PKT-1 and PKT-2) a purification step between end repair with PKT and treatment with Taq DNA pol at 65 °C for 30 min. ( d ) Comparison of library yield of the three sets described above with or without (PKT on the left) a purification step between end repair and 3′ A-tailing indicates that purification caused substantial loss of library DNA.

    Article Snippet: Enzyme mix PKT was comprised of approximately 1,200 units/ml T4 DNA polymerase, 2,000 units/ml T4 PNK and 2,000 units/ml Taq DNA polymerase (NEB) while PK contained T4 DNA polymerase and T4 PNK only.

    Techniques: Polymerase Chain Reaction, Sequencing, Generated, Produced, Purification, Incubation

    Enzyme immobilization and comparison of Illumina library preparation protocols. ( a ) A schematic of covalent conjugation of SNAP-tagged enzyme fusion proteins to magnetic beads functionalized with O 6 . ( b ) Workflow for library construction using immobilized enzymes for Illumina sequencing. A typical streamlined protocol for Illumina library construction is modified by employing immobilized enzymes to catalyze end repair and 3′ A-tailing. This method utilizes SNAP-tagged T4 DNA pol and PNK covalently conjugated to BG-functionalized magnetic beads to carry out end repair of fragmented DNA at 20°C (or 37 °C) for 30 min. The enzymes are removed by magnetic separation from the DNA pool, which is subjected to 3′ A-tailing at 37 °C for 30 min using immobilized Taq DNA pol. ( c ) Streamlined protocol for Illumina amplification-free library preparation using soluble enzymes. Typically, end repair and 3′ A-tailing of fragmented DNA are catalyzed by an enzyme mixture at 20 °C for 30 min, followed by heat treatment at 65 °C for 30 min. ( d ) The workflow of Illumina TruSeq DNA PCR-free LT Library Prep Kit with a purification step. End repair is performed for 30 min at 30 °C, followed by a bead-based step for clean up and size selection. 3′ A-tailing is carried out for 30 min at 37 °C with a subsequent treatment for 5 min at 70 °C. Each library was ligated to preannealed full-length paired-end Illumina adaptors, size-selected and analyzed, and sequenced on an Illumina sequencing platform.

    Journal: Scientific Reports

    Article Title: Solid-phase enzyme catalysis of DNA end repair and 3′ A-tailing reduces GC-bias in next-generation sequencing of human genomic DNA

    doi: 10.1038/s41598-018-34079-2

    Figure Lengend Snippet: Enzyme immobilization and comparison of Illumina library preparation protocols. ( a ) A schematic of covalent conjugation of SNAP-tagged enzyme fusion proteins to magnetic beads functionalized with O 6 . ( b ) Workflow for library construction using immobilized enzymes for Illumina sequencing. A typical streamlined protocol for Illumina library construction is modified by employing immobilized enzymes to catalyze end repair and 3′ A-tailing. This method utilizes SNAP-tagged T4 DNA pol and PNK covalently conjugated to BG-functionalized magnetic beads to carry out end repair of fragmented DNA at 20°C (or 37 °C) for 30 min. The enzymes are removed by magnetic separation from the DNA pool, which is subjected to 3′ A-tailing at 37 °C for 30 min using immobilized Taq DNA pol. ( c ) Streamlined protocol for Illumina amplification-free library preparation using soluble enzymes. Typically, end repair and 3′ A-tailing of fragmented DNA are catalyzed by an enzyme mixture at 20 °C for 30 min, followed by heat treatment at 65 °C for 30 min. ( d ) The workflow of Illumina TruSeq DNA PCR-free LT Library Prep Kit with a purification step. End repair is performed for 30 min at 30 °C, followed by a bead-based step for clean up and size selection. 3′ A-tailing is carried out for 30 min at 37 °C with a subsequent treatment for 5 min at 70 °C. Each library was ligated to preannealed full-length paired-end Illumina adaptors, size-selected and analyzed, and sequenced on an Illumina sequencing platform.

    Article Snippet: Enzyme mix PKT was comprised of approximately 1,200 units/ml T4 DNA polymerase, 2,000 units/ml T4 PNK and 2,000 units/ml Taq DNA polymerase (NEB) while PK contained T4 DNA polymerase and T4 PNK only.

    Techniques: Conjugation Assay, Magnetic Beads, Sequencing, Modification, Amplification, Polymerase Chain Reaction, Purification, Selection

    DNA polymerization by PCR using a caged primer (17 nt) containing the first caged thymidine (blue square) 10 nt from the 5′ end. PCR generates a caged template which results in a stop of Taq and Pfu polymerase due to the presence of a single-caged thymidine.

    Journal: Nucleic Acids Research

    Article Title: Restriction enzyme-free mutagenesis via the light regulation of DNA polymerization

    doi: 10.1093/nar/gkp150

    Figure Lengend Snippet: DNA polymerization by PCR using a caged primer (17 nt) containing the first caged thymidine (blue square) 10 nt from the 5′ end. PCR generates a caged template which results in a stop of Taq and Pfu polymerase due to the presence of a single-caged thymidine.

    Article Snippet: These results demonstrate that both Taq DNA polymerase and Phusion DNA polymerase are stopped by a single-caged thymidine.

    Techniques: Polymerase Chain Reaction

    Effects of a caged thymidine nucleobase on the PCR. ( A ) PCR product using Phusion DNA Polymerase. Polymerization is halted in the presence of a caging group, which is confirmed using the truncated primer P3 that affords the same length product. ( B ) PCR product using Taq DNA polymerase. Polymerization is halted in the presence of a caging group, which is confirmed using the truncated primer P3 that affords the same length product.

    Journal: Nucleic Acids Research

    Article Title: Restriction enzyme-free mutagenesis via the light regulation of DNA polymerization

    doi: 10.1093/nar/gkp150

    Figure Lengend Snippet: Effects of a caged thymidine nucleobase on the PCR. ( A ) PCR product using Phusion DNA Polymerase. Polymerization is halted in the presence of a caging group, which is confirmed using the truncated primer P3 that affords the same length product. ( B ) PCR product using Taq DNA polymerase. Polymerization is halted in the presence of a caging group, which is confirmed using the truncated primer P3 that affords the same length product.

    Article Snippet: These results demonstrate that both Taq DNA polymerase and Phusion DNA polymerase are stopped by a single-caged thymidine.

    Techniques: Polymerase Chain Reaction