user enzyme  (New England Biolabs)


Bioz Verified Symbol New England Biolabs is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Name:
    USER Enzyme
    Description:
    USER Enzyme 250 units
    Catalog Number:
    m5505l
    Price:
    292
    Size:
    250 units
    Category:
    Other Enzymes
    Buy from Supplier


    Structured Review

    New England Biolabs user enzyme
    USER Enzyme
    USER Enzyme 250 units
    https://www.bioz.com/result/user enzyme/product/New England Biolabs
    Average 90 stars, based on 405 article reviews
    Price from $9.99 to $1999.99
    user enzyme - by Bioz Stars, 2020-01
    90/100 stars

    Images

    1) Product Images from "A versatile element for gene addition in bacterial chromosomes"

    Article Title: A versatile element for gene addition in bacterial chromosomes

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkr1085

    Design of primers for USER cloning with pMS26. Two choices of translation signal in the 5′-UTR are shown. The gene-specific sequence illustrated is of fnuDIIM . The underlined 21 bp of longer translation signal (LTS) is from the tacp regulatory region ( 34 ) ; the short signal (STS) is a truncation of it. The LTS and downstream primers illustrated are the same as primers 5 and 6 of Table 4 ; the STS construct was made but not used in this report. Fusion of lacZ to the signal as shown creates an RBS/ATG spacing of six, within the usual range of spacing ( 35 ); lacZ native spacing is seven ( 36 ).
    Figure Legend Snippet: Design of primers for USER cloning with pMS26. Two choices of translation signal in the 5′-UTR are shown. The gene-specific sequence illustrated is of fnuDIIM . The underlined 21 bp of longer translation signal (LTS) is from the tacp regulatory region ( 34 ) ; the short signal (STS) is a truncation of it. The LTS and downstream primers illustrated are the same as primers 5 and 6 of Table 4 ; the STS construct was made but not used in this report. Fusion of lacZ to the signal as shown creates an RBS/ATG spacing of six, within the usual range of spacing ( 35 ); lacZ native spacing is seven ( 36 ).

    Techniques Used: Clone Assay, Sequencing, Construct

    Related Articles

    Diagnostic Assay:

    Article Title: Who Needs Neighbors? PKS8 Is a Stand-Alone Gene in Fusarium graminearum Responsible for Production of Gibepyrones and Prolipyrone B
    Article Snippet: The PCR fragments were cloned into a linearized pRF-HU2E vector under control of the gpdA promoter by a four fragment cloning step using the USER enzyme™ (New England Biolabs, Ipswich, MA, USA) and verified by colony PCR [ ]. .. Transformation of F. graminearum was carried out by Agrobacterium tumefaciens mediated transformation as described previously [ ] and the resulting mutants were verified by diagnostic PCR using a forward primer annealing to gDNA outside the border region and a reverse primer annealing to the hygromycin resistance gene.

    Clone Assay:

    Article Title: Who Needs Neighbors? PKS8 Is a Stand-Alone Gene in Fusarium graminearum Responsible for Production of Gibepyrones and Prolipyrone B
    Article Snippet: .. The PCR fragments were cloned into a linearized pRF-HU2E vector under control of the gpdA promoter by a four fragment cloning step using the USER enzyme™ (New England Biolabs, Ipswich, MA, USA) and verified by colony PCR [ ]. .. Transformation of F. graminearum was carried out by Agrobacterium tumefaciens mediated transformation as described previously [ ] and the resulting mutants were verified by diagnostic PCR using a forward primer annealing to gDNA outside the border region and a reverse primer annealing to the hygromycin resistance gene.

    Article Title: Fluorescently Labeled DNA Interacts with Competence and Recombination Proteins and Is Integrated and Expressed Following Natural Transformation of Bacillus subtilis
    Article Snippet: The B. subtilis 168 amyE ::Pxyl -comK-cm comFC-gfp-tet strain was obtained by USER cloning. .. The different components of the construct were obtained by PCR with PfuX7 , treated with USER enzyme (NEB), ligated overnight at 4°C, and transformed directly into B. subtilis 168 amyE ::Pxyl -comK-cm for integration into the native locus.

    Article Title: Continuous directed evolution of aminoacyl-tRNA synthetases
    Article Snippet: .. Plasmids and selection phage were prepared using isothermal assembly with Gibson Assembly 2x Master Mix (New England Biolabs), USER cloning with USER enzyme (New England Biolabs), or ligation cycling reaction with Ampligase (Epicentre). .. Genes were either synthesized from gBlock gene fragments (Integrated DNA Technologies) or PCR amplified from native sources.

    Amplification:

    Article Title: Who Needs Neighbors? PKS8 Is a Stand-Alone Gene in Fusarium graminearum Responsible for Production of Gibepyrones and Prolipyrone B
    Article Snippet: Overexpression of PKS8 in F. graminearum The flanking regions of PKS8 was PCR amplified from F. graminearum PH-1 (NRRL31084) using primers PKS8 -O1 to PKS8 -O4 listed in and Pfu polymerase (Stratagene, La Jolla, CA, USA). .. The PCR fragments were cloned into a linearized pRF-HU2E vector under control of the gpdA promoter by a four fragment cloning step using the USER enzyme™ (New England Biolabs, Ipswich, MA, USA) and verified by colony PCR [ ].

    Article Title: Fluorescently Labeled DNA Interacts with Competence and Recombination Proteins and Is Integrated and Expressed Following Natural Transformation of Bacillus subtilis
    Article Snippet: The different components of the construct were obtained by PCR with PfuX7 , treated with USER enzyme (NEB), ligated overnight at 4°C, and transformed directly into B. subtilis 168 amyE ::Pxyl -comK-cm for integration into the native locus. .. B. subtilis 168 amyE ::Pxyl -comK thrC ::Pspank -parB-gfp and B. subtilis 168 amyE ::Pxyl -comK thrC ::Pspank -parB-mkate2 were created by amplification of parB-mkate2 from pMK17 and parB-gfp from pMK11 ( , , ) with primers 133 and 134. parB-gfp and parB-mkate were cloned into pMB002 using NheI and HindIII (FastDigest; Thermo Scientific), ligated with T4 ligase (Thermo Scientific), transformed into Escherichia coli DH5α, and sequenced.

    Article Title: Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
    Article Snippet: Homologous gene sequences were then amplified with PCR using a Pfu Turbo Cx Hotstart DNA polymerase (Agilent Technologies, USA) and the following settings: 95°C for 2 minutes, 30 cycles: 95°C for 30s, 55°C for 30s, 72°C for 1 minute; 75°C for 10 minutes. .. PCR products were purified with Illustra GFX PCR DNA and a Gel Band Purification Kit (GE Healthcare, UK) and ligated to linearized vector pRF-HU2 with USER enzyme (New England BioLabs, USA).

    Article Title: Continuous directed evolution of aminoacyl-tRNA synthetases
    Article Snippet: Plasmids and selection phage were prepared using isothermal assembly with Gibson Assembly 2x Master Mix (New England Biolabs), USER cloning with USER enzyme (New England Biolabs), or ligation cycling reaction with Ampligase (Epicentre). .. Genes were either synthesized from gBlock gene fragments (Integrated DNA Technologies) or PCR amplified from native sources.

    Whole Genome Amplification:

    Article Title: Mycobacterium leprae genomes from a British medieval leprosy hospital: towards understanding an ancient epidemic
    Article Snippet: Paragraph title: Whole genome amplification, enrichment and analysis ... Both involved removing uracil residues with USER enzyme (New England Biolabs, Hitchin, UK), repairing the DNA, ligating linkers to the ends, and PCR amplifying before enriching for M. leprae sequences using microarrays (once at the University of Tübingen and twice at the University of Surrey).

    Synthesized:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: Immediately after, a second strand cDNA was synthesized using DNA polymerase I and RNase H. After adenylation reaction of 3′ ends of DNA fragments, NEBNext Adaptor containing hairpin loop structure were ligated for hybridization. .. The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C.

    Article Title: Transcriptome and gene expression analysis of DHA producer Aurantiochytrium under low temperature conditions
    Article Snippet: First strand cDNA was synthesized using random hexamer primers and M-MuLV Reverse Transcriptase (RNase H− ). .. Then, 3 μL of USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min, followed by 5 min at 95 °C before PCR.

    Article Title: Screening and characterization of long noncoding RNAs involved in the albinism of Ananas comosus var. bracteatus leaves
    Article Snippet: First-strand cDNA was synthesized using a random hexamer primer and reverse transcriptase. .. Then, 3 μl of USER enzyme (NEB, USA) was incubated with the size-selected, adaptor-ligated cDNA at 37°C for 15 min before PCR.

    Article Title: Comparative Transcriptome Analysis of Genes Involved in Anthocyanin Biosynthesis in Red and Green Walnut (Juglans regia L.)
    Article Snippet: First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase. .. Then 3 μL USER Enzyme (NEB) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR.

    Article Title: Transcriptional Responses of Creeping Bentgrass to 2,3-Butanediol, a Bacterial Volatile Compound (BVC) Analogue
    Article Snippet: First strand cDNA was synthesized using a random hexamer primer and M-MuLV reverse transcriptase (RNase H-). .. Then 3 μL USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR.

    Article Title: Continuous directed evolution of aminoacyl-tRNA synthetases
    Article Snippet: Plasmids and selection phage were prepared using isothermal assembly with Gibson Assembly 2x Master Mix (New England Biolabs), USER cloning with USER enzyme (New England Biolabs), or ligation cycling reaction with Ampligase (Epicentre). .. Genes were either synthesized from gBlock gene fragments (Integrated DNA Technologies) or PCR amplified from native sources.

    Article Title: An influential meal: host plant dependent transcriptional variation in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae)
    Article Snippet: Synthesis of first strand cDNA was performed using random hexamer primers and M-MuLV Reverse Transcriptase (RNase H− ), subsequently second strand cDNA was synthesized using DNA polymerase I and RNase H. After terminal repair of the sequences, the 3′ ends were adenylated and NEBnext adapters (NEB, MA, USA) were ligated. .. To the size-selected cDNA, 3 μl USER Enzyme (NEB, MA, USA) was added and heated to 37 °C for 15 min followed by 5 min at 95 °C.

    Construct:

    Article Title: Fluorescently Labeled DNA Interacts with Competence and Recombination Proteins and Is Integrated and Expressed Following Natural Transformation of Bacillus subtilis
    Article Snippet: .. The different components of the construct were obtained by PCR with PfuX7 , treated with USER enzyme (NEB), ligated overnight at 4°C, and transformed directly into B. subtilis 168 amyE ::Pxyl -comK-cm for integration into the native locus. ..

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: Sequencing libraries were constructed using NEBNext® Ultra™ RNA Library Prep Kit for Illumina® (New England Biolabs, Ipswich, MA, USA). .. The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C.

    Article Title: Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
    Article Snippet: PCR products were purified with Illustra GFX PCR DNA and a Gel Band Purification Kit (GE Healthcare, UK) and ligated to linearized vector pRF-HU2 with USER enzyme (New England BioLabs, USA). .. Vector constructs were multiplied in E . coli DH5α cells and isolated with a High Pure Plasmid Isolation Kit (Roche, Life Science, USA).

    Real-time Polymerase Chain Reaction:

    Article Title: Screening and characterization of long noncoding RNAs involved in the albinism of Ananas comosus var. bracteatus leaves
    Article Snippet: Then, 3 μl of USER enzyme (NEB, USA) was incubated with the size-selected, adaptor-ligated cDNA at 37°C for 15 min before PCR. .. Finally, the PCR products were purified (AMPure XP system), and library quality was assessed on an Agilent Bioanalyzer 2100 and by qPCR.

    Random Hexamer Labeling:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: First, strand cDNA was produced based on the random hexamer primer and M-MuLV Reverse Transcriptase. .. The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C.

    Article Title: Transcriptome and gene expression analysis of DHA producer Aurantiochytrium under low temperature conditions
    Article Snippet: First strand cDNA was synthesized using random hexamer primers and M-MuLV Reverse Transcriptase (RNase H− ). .. Then, 3 μL of USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min, followed by 5 min at 95 °C before PCR.

    Article Title: Metabolomic and Transcriptomic Analyses Reveal That a MADS-Box Transcription Factor TDR4 Regulates Tomato Fruit Quality
    Article Snippet: These fragments were used to synthesize first-strand cDNA using random hexamer primers and M-MuLV Reverse Transcriptase. .. USER Enzyme (New England Biolabs) was subsequently used with size-selected, adaptor-ligated cDNA.

    Article Title: Screening and characterization of long noncoding RNAs involved in the albinism of Ananas comosus var. bracteatus leaves
    Article Snippet: First-strand cDNA was synthesized using a random hexamer primer and reverse transcriptase. .. Then, 3 μl of USER enzyme (NEB, USA) was incubated with the size-selected, adaptor-ligated cDNA at 37°C for 15 min before PCR.

    Article Title: Comparative Transcriptome Analysis of Genes Involved in Anthocyanin Biosynthesis in Red and Green Walnut (Juglans regia L.)
    Article Snippet: First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse Transcriptase. .. Then 3 μL USER Enzyme (NEB) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR.

    Article Title: Transcriptional Responses of Creeping Bentgrass to 2,3-Butanediol, a Bacterial Volatile Compound (BVC) Analogue
    Article Snippet: First strand cDNA was synthesized using a random hexamer primer and M-MuLV reverse transcriptase (RNase H-). .. Then 3 μL USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR.

    Article Title: An influential meal: host plant dependent transcriptional variation in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae)
    Article Snippet: Synthesis of first strand cDNA was performed using random hexamer primers and M-MuLV Reverse Transcriptase (RNase H− ), subsequently second strand cDNA was synthesized using DNA polymerase I and RNase H. After terminal repair of the sequences, the 3′ ends were adenylated and NEBnext adapters (NEB, MA, USA) were ligated. .. To the size-selected cDNA, 3 μl USER Enzyme (NEB, MA, USA) was added and heated to 37 °C for 15 min followed by 5 min at 95 °C.

    Expressing:

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C. .. RPKM (reads per kilobase of gene length per million reads) values for RefSeq genes were computed using tag counting scripts and used to analyse the expression level of genes.

    Transformation Assay:

    Article Title: Who Needs Neighbors? PKS8 Is a Stand-Alone Gene in Fusarium graminearum Responsible for Production of Gibepyrones and Prolipyrone B
    Article Snippet: The PCR fragments were cloned into a linearized pRF-HU2E vector under control of the gpdA promoter by a four fragment cloning step using the USER enzyme™ (New England Biolabs, Ipswich, MA, USA) and verified by colony PCR [ ]. .. Transformation of F. graminearum was carried out by Agrobacterium tumefaciens mediated transformation as described previously [ ] and the resulting mutants were verified by diagnostic PCR using a forward primer annealing to gDNA outside the border region and a reverse primer annealing to the hygromycin resistance gene.

    Article Title: Fluorescently Labeled DNA Interacts with Competence and Recombination Proteins and Is Integrated and Expressed Following Natural Transformation of Bacillus subtilis
    Article Snippet: .. The different components of the construct were obtained by PCR with PfuX7 , treated with USER enzyme (NEB), ligated overnight at 4°C, and transformed directly into B. subtilis 168 amyE ::Pxyl -comK-cm for integration into the native locus. ..

    Article Title: Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
    Article Snippet: PCR products were purified with Illustra GFX PCR DNA and a Gel Band Purification Kit (GE Healthcare, UK) and ligated to linearized vector pRF-HU2 with USER enzyme (New England BioLabs, USA). .. V . nonalfalfae knockout mutants were generated with Agrobacterium tumefaciens mediated transformation (ATMT) [ ] using acetosyringone (Sigma Aldrich, USA).

    Over Expression:

    Article Title: Who Needs Neighbors? PKS8 Is a Stand-Alone Gene in Fusarium graminearum Responsible for Production of Gibepyrones and Prolipyrone B
    Article Snippet: Paragraph title: 3.2. Overexpression of PKS8 in F. graminearum ... The PCR fragments were cloned into a linearized pRF-HU2E vector under control of the gpdA promoter by a four fragment cloning step using the USER enzyme™ (New England Biolabs, Ipswich, MA, USA) and verified by colony PCR [ ].

    Hybridization:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: Immediately after, a second strand cDNA was synthesized using DNA polymerase I and RNase H. After adenylation reaction of 3′ ends of DNA fragments, NEBNext Adaptor containing hairpin loop structure were ligated for hybridization. .. The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C.

    Article Title: Transcriptome and gene expression analysis of DHA producer Aurantiochytrium under low temperature conditions
    Article Snippet: After the 3′ ends of the DNA fragments were adenylated, NEBNext adaptors with a hairpin loop structure were ligated to prepare for hybridization. .. Then, 3 μL of USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min, followed by 5 min at 95 °C before PCR.

    Article Title: Screening and characterization of long noncoding RNAs involved in the albinism of Ananas comosus var. bracteatus leaves
    Article Snippet: After adenylation of the 3' ends of the DNA fragments, NEBNext adaptors with a hairpin loop structure were ligated to prepare each sample for hybridization. .. Then, 3 μl of USER enzyme (NEB, USA) was incubated with the size-selected, adaptor-ligated cDNA at 37°C for 15 min before PCR.

    Article Title: Comparative Transcriptome Analysis of Genes Involved in Anthocyanin Biosynthesis in Red and Green Walnut (Juglans regia L.)
    Article Snippet: After adenylation of 3′ ends of DNA fragments, NEBNext Adaptor with hairpin loop structure were ligated to prepare for hybridization. .. Then 3 μL USER Enzyme (NEB) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR.

    Article Title: Transcriptional Responses of Creeping Bentgrass to 2,3-Butanediol, a Bacterial Volatile Compound (BVC) Analogue
    Article Snippet: After adenylation of 3′ ends of DNA fragments, NEBNext adaptors with a hairpin loop structure were ligated to prepare for hybridization. .. Then 3 μL USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR.

    Ligation:

    Article Title: Continuous directed evolution of aminoacyl-tRNA synthetases
    Article Snippet: .. Plasmids and selection phage were prepared using isothermal assembly with Gibson Assembly 2x Master Mix (New England Biolabs), USER cloning with USER enzyme (New England Biolabs), or ligation cycling reaction with Ampligase (Epicentre). .. Genes were either synthesized from gBlock gene fragments (Integrated DNA Technologies) or PCR amplified from native sources.

    Generated:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C. .. Finally, PCR products were purified by AMPure XP system and the Agilent Bioanalyzer 2100 system was used to assess sequencing library quality and the clusters were sequenced on an Illumina Hiseq 2000 platform and paired-end reads were generated.

    Article Title: Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
    Article Snippet: PCR products were purified with Illustra GFX PCR DNA and a Gel Band Purification Kit (GE Healthcare, UK) and ligated to linearized vector pRF-HU2 with USER enzyme (New England BioLabs, USA). .. V . nonalfalfae knockout mutants were generated with Agrobacterium tumefaciens mediated transformation (ATMT) [ ] using acetosyringone (Sigma Aldrich, USA).

    Article Title: Mitogenome evolution in the last surviving woolly mammoth population reveals neutral and functional consequences of small population size
    Article Snippet: DATA PREPARATION Sample E469D was extracted using a method optimized for highly degraded samples (Dabney et al. ), while sequence data for samples labeled “E” come from (Palkopoulou et al. ), but with new consensus sequences generated in this study. .. Double stranded Illumina libraries were prepared from 20 μL of DNA extract according to Meyer and Kircher , using uracil‐treatment with the USER enzyme (New England Biolabs; Briggs et al. ).

    Polymerase Chain Reaction:

    Article Title: Who Needs Neighbors? PKS8 Is a Stand-Alone Gene in Fusarium graminearum Responsible for Production of Gibepyrones and Prolipyrone B
    Article Snippet: .. The PCR fragments were cloned into a linearized pRF-HU2E vector under control of the gpdA promoter by a four fragment cloning step using the USER enzyme™ (New England Biolabs, Ipswich, MA, USA) and verified by colony PCR [ ]. .. Transformation of F. graminearum was carried out by Agrobacterium tumefaciens mediated transformation as described previously [ ] and the resulting mutants were verified by diagnostic PCR using a forward primer annealing to gDNA outside the border region and a reverse primer annealing to the hygromycin resistance gene.

    Article Title: Fluorescently Labeled DNA Interacts with Competence and Recombination Proteins and Is Integrated and Expressed Following Natural Transformation of Bacillus subtilis
    Article Snippet: .. The different components of the construct were obtained by PCR with PfuX7 , treated with USER enzyme (NEB), ligated overnight at 4°C, and transformed directly into B. subtilis 168 amyE ::Pxyl -comK-cm for integration into the native locus. ..

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C. .. Then PCR reaction was performed using Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer.

    Article Title: Transcriptome and gene expression analysis of DHA producer Aurantiochytrium under low temperature conditions
    Article Snippet: .. Then, 3 μL of USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min, followed by 5 min at 95 °C before PCR. .. Then, PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers, and an index (X) primer.

    Article Title: Integrative Analyses of Long Non-coding RNA and mRNA Involved in Piglet Ileum Immune Response to Clostridium perfringens Type C Infection
    Article Snippet: .. Then 3 μL USER Enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37°C for 15 min followed by 5 min at 95°C before PCR, then PCR reaction was performed using Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer. .. At last, products were purified (AMPure XP system) and library quality were assessed on the Agilent Bioanalyzer 2100 system.

    Article Title: Metabolomic and Transcriptomic Analyses Reveal That a MADS-Box Transcription Factor TDR4 Regulates Tomato Fruit Quality
    Article Snippet: USER Enzyme (New England Biolabs) was subsequently used with size-selected, adaptor-ligated cDNA. .. Then, PCR was carried out with Phusion High-Fidelity DNA polymerase, universal PCR primers, and Index (X) Primer.

    Article Title: Mycobacterium leprae genomes from a British medieval leprosy hospital: towards understanding an ancient epidemic
    Article Snippet: .. Both involved removing uracil residues with USER enzyme (New England Biolabs, Hitchin, UK), repairing the DNA, ligating linkers to the ends, and PCR amplifying before enriching for M. leprae sequences using microarrays (once at the University of Tübingen and twice at the University of Surrey). ..

    Article Title: Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
    Article Snippet: .. PCR products were purified with Illustra GFX PCR DNA and a Gel Band Purification Kit (GE Healthcare, UK) and ligated to linearized vector pRF-HU2 with USER enzyme (New England BioLabs, USA). .. Vector constructs were multiplied in E . coli DH5α cells and isolated with a High Pure Plasmid Isolation Kit (Roche, Life Science, USA).

    Article Title: Screening and characterization of long noncoding RNAs involved in the albinism of Ananas comosus var. bracteatus leaves
    Article Snippet: .. Then, 3 μl of USER enzyme (NEB, USA) was incubated with the size-selected, adaptor-ligated cDNA at 37°C for 15 min before PCR. .. PCR was then performed with Phusion high-fidelity DNA polymerase, universal PCR primers and the index (X) primer.

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: .. Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C. .. The 300-bp libraries were used for cluster generation on a HiSeq 2000 (Illumina).

    Article Title: Transcriptional Responses of Creeping Bentgrass to 2,3-Butanediol, a Bacterial Volatile Compound (BVC) Analogue
    Article Snippet: .. Then 3 μL USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR. .. Then PCR was performed with Phusion High-Fidelity DNA polymerase, universal PCR primers and index (X) primer.

    Article Title: Continuous directed evolution of aminoacyl-tRNA synthetases
    Article Snippet: PCR was performed with Q5 Hot Start High-Fidelity DNA polymerase (New England Biolabs) when unmodified primers were used, and Phusion U Hot Start DNA polymerase (Thermo Fisher Scientific) was used when deoxyuridine-containing primers were required for USER cloning. .. Plasmids and selection phage were prepared using isothermal assembly with Gibson Assembly 2x Master Mix (New England Biolabs), USER cloning with USER enzyme (New England Biolabs), or ligation cycling reaction with Ampligase (Epicentre).

    Article Title: An influential meal: host plant dependent transcriptional variation in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae)
    Article Snippet: To the size-selected cDNA, 3 μl USER Enzyme (NEB, MA, USA) was added and heated to 37 °C for 15 min followed by 5 min at 95 °C. .. The cDNA was enriched by PCR reactions using Phusion High-Fidelity DNA polymerase, universal PCR primers and index primers.

    RNA Sequencing Assay:

    Article Title: Integrative Analyses of Long Non-coding RNA and mRNA Involved in Piglet Ileum Immune Response to Clostridium perfringens Type C Infection
    Article Snippet: Paragraph title: Library Preparation and RNA-Seq Data Acquisition ... Then 3 μL USER Enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37°C for 15 min followed by 5 min at 95°C before PCR, then PCR reaction was performed using Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer.

    Article Title: Metabolomic and Transcriptomic Analyses Reveal That a MADS-Box Transcription Factor TDR4 Regulates Tomato Fruit Quality
    Article Snippet: Paragraph title: RNA-Seq and Data Processing ... USER Enzyme (New England Biolabs) was subsequently used with size-selected, adaptor-ligated cDNA.

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C. .. RNA-Seq reads were uniquely mapped to the human (hg19) and mouse (mm9) reference genomes using the Eland or BWA program, allowing 1 mismatch, and subsequently used for bioinformatic analysis.

    Article Title: An influential meal: host plant dependent transcriptional variation in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae)
    Article Snippet: Paragraph title: RNA sequencing ... To the size-selected cDNA, 3 μl USER Enzyme (NEB, MA, USA) was added and heated to 37 °C for 15 min followed by 5 min at 95 °C.

    Magnetic Beads:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: Briefly, purified mRNA was obtained from total RNA (1 µg) per sample using poly-T oligo-attached magnetic beads; then, the mRNA was fragmented using divalent cations under elevated temperature in NEBNext First Strand Synthesis Reaction Buffer (5X). .. The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C.

    Article Title: Metabolomic and Transcriptomic Analyses Reveal That a MADS-Box Transcription Factor TDR4 Regulates Tomato Fruit Quality
    Article Snippet: Briefly, mRNA was enriched using oligo (dT)-attached magnetic beads. .. USER Enzyme (New England Biolabs) was subsequently used with size-selected, adaptor-ligated cDNA.

    Article Title: An influential meal: host plant dependent transcriptional variation in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae)
    Article Snippet: The mRNA was purified using poly-T oligo-attached magnetic beads followed by fragmentation using NEBNext First Strand Synthesis Reaction Buffer (5x). .. To the size-selected cDNA, 3 μl USER Enzyme (NEB, MA, USA) was added and heated to 37 °C for 15 min followed by 5 min at 95 °C.

    Mutagenesis:

    Article Title: Continuous directed evolution of aminoacyl-tRNA synthetases
    Article Snippet: Plasmids and selection phage were prepared using isothermal assembly with Gibson Assembly 2x Master Mix (New England Biolabs), USER cloning with USER enzyme (New England Biolabs), or ligation cycling reaction with Ampligase (Epicentre). .. Premature stop codons and single point mutations were placed into genes using the Q5 Site-Directed Mutagenesis kit (New England Biolabs).

    Isolation:

    Article Title: Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
    Article Snippet: PCR products were purified with Illustra GFX PCR DNA and a Gel Band Purification Kit (GE Healthcare, UK) and ligated to linearized vector pRF-HU2 with USER enzyme (New England BioLabs, USA). .. Vector constructs were multiplied in E . coli DH5α cells and isolated with a High Pure Plasmid Isolation Kit (Roche, Life Science, USA).

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: After washing in Ca2+ - and Mg2+ -containing PBS, RNA was isolated and purified using the RNAeasy kit (Qiagen), followed by DNase treatment (Qiagen). .. Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C.

    Purification:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: .. The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C. .. Then PCR reaction was performed using Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer.

    Article Title: Transcriptome and gene expression analysis of DHA producer Aurantiochytrium under low temperature conditions
    Article Snippet: To preferentially select cDNA fragments of 150–200 bp in length, the library fragments were purified using the AMPure XP system (Beckman Coulter, Beverly, USA). .. Then, 3 μL of USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min, followed by 5 min at 95 °C before PCR.

    Article Title: Integrative Analyses of Long Non-coding RNA and mRNA Involved in Piglet Ileum Immune Response to Clostridium perfringens Type C Infection
    Article Snippet: To select cDNA fragments of preferentially 150–200 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA). .. Then 3 μL USER Enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37°C for 15 min followed by 5 min at 95°C before PCR, then PCR reaction was performed using Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer.

    Article Title: Metabolomic and Transcriptomic Analyses Reveal That a MADS-Box Transcription Factor TDR4 Regulates Tomato Fruit Quality
    Article Snippet: In order to select cDNA fragments of the appropriate size, library fragments were purified with the AMPure XP system (Beckman Coulter, Beverly, MA, United States). .. USER Enzyme (New England Biolabs) was subsequently used with size-selected, adaptor-ligated cDNA.

    Article Title: Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
    Article Snippet: .. PCR products were purified with Illustra GFX PCR DNA and a Gel Band Purification Kit (GE Healthcare, UK) and ligated to linearized vector pRF-HU2 with USER enzyme (New England BioLabs, USA). .. Vector constructs were multiplied in E . coli DH5α cells and isolated with a High Pure Plasmid Isolation Kit (Roche, Life Science, USA).

    Article Title: Screening and characterization of long noncoding RNAs involved in the albinism of Ananas comosus var. bracteatus leaves
    Article Snippet: To preferentially select insert fragments that were 150~200 bp in length, the library fragments were purified with AMPure XP beads (Beckman Coulter, Beverly, USA). .. Then, 3 μl of USER enzyme (NEB, USA) was incubated with the size-selected, adaptor-ligated cDNA at 37°C for 15 min before PCR.

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: Second-strand cDNA was synthesised by adding 91.8 μl H2 O, 5 μg random hexamers, 4 μl 5× first-strand buffer, 2 μl 100 mM DTT, 4 μl 10 mM dNTPs with dTTP replaced by dUTP, 30 μl 5× second-strand buffer, 40 U E. coli DNA polymerase, 10 U E . coli DNA ligase and 2 U E . coli RNase H, and incubated at 16 °C for 2 h, followed by incubation with 10 U T4 polymerase at 16 °C for 10 min. Double-stranded cDNA was purified using a Qiagen MinElute column and used for Illumina sample preparation and sequencing according to the Illumina protocol. .. Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C.

    Article Title: Transcriptional Responses of Creeping Bentgrass to 2,3-Butanediol, a Bacterial Volatile Compound (BVC) Analogue
    Article Snippet: In order to select cDNA fragments of preferentially 150~200 bp in length, the library fragments were purified with the AMPure XP system (Beckman Coulter, Beverly, MA, USA). .. Then 3 μL USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR.

    Article Title: Continuous directed evolution of aminoacyl-tRNA synthetases
    Article Snippet: In all other experiments, water was purified by a MilliQ purification system (EMD Millipore). .. Plasmids and selection phage were prepared using isothermal assembly with Gibson Assembly 2x Master Mix (New England Biolabs), USER cloning with USER enzyme (New England Biolabs), or ligation cycling reaction with Ampligase (Epicentre).

    Article Title: An influential meal: host plant dependent transcriptional variation in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae)
    Article Snippet: The library fragments were purified, for a 150–200 bp size range, with AMpure XP purification kit (Beckman Coulter, MA, USA). .. To the size-selected cDNA, 3 μl USER Enzyme (NEB, MA, USA) was added and heated to 37 °C for 15 min followed by 5 min at 95 °C.

    Sequencing:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: Paragraph title: 2.2. RNA Extraction, Library Preparation, Sequencing and De Novo Assembly ... The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C.

    Article Title: Transcriptome and gene expression analysis of DHA producer Aurantiochytrium under low temperature conditions
    Article Snippet: Paragraph title: RNA extraction, library preparation and sequencing ... Then, 3 μL of USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min, followed by 5 min at 95 °C before PCR.

    Article Title: Screening and characterization of long noncoding RNAs involved in the albinism of Ananas comosus var. bracteatus leaves
    Article Snippet: Paragraph title: LncRNA library construction and sequencing ... Then, 3 μl of USER enzyme (NEB, USA) was incubated with the size-selected, adaptor-ligated cDNA at 37°C for 15 min before PCR.

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: Paragraph title: mRNA sequencing ... Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C.

    Article Title: Comparative Transcriptome Analysis of Genes Involved in Anthocyanin Biosynthesis in Red and Green Walnut (Juglans regia L.)
    Article Snippet: Paragraph title: 4.3. Library Preparation for Transcriptome Sequencing ... Then 3 μL USER Enzyme (NEB) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR.

    Article Title: Mitogenome evolution in the last surviving woolly mammoth population reveals neutral and functional consequences of small population size
    Article Snippet: DATA PREPARATION Sample E469D was extracted using a method optimized for highly degraded samples (Dabney et al. ), while sequence data for samples labeled “E” come from (Palkopoulou et al. ), but with new consensus sequences generated in this study. .. Double stranded Illumina libraries were prepared from 20 μL of DNA extract according to Meyer and Kircher , using uracil‐treatment with the USER enzyme (New England Biolabs; Briggs et al. ).

    Article Title: Transcriptional Responses of Creeping Bentgrass to 2,3-Butanediol, a Bacterial Volatile Compound (BVC) Analogue
    Article Snippet: Paragraph title: 4.5. Library Preparation and Transcriptome Sequencing ... Then 3 μL USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min followed by 5 min at 95 °C before PCR.

    Article Title: An influential meal: host plant dependent transcriptional variation in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae)
    Article Snippet: The NEBNext Ultra RNA Library Prep Kit for Illumina sequencing (NEB, MA, USA) following the protocol as provided by the manufacturer was used for library preparation, index sequences were added to trace back the sequences to each sample. .. To the size-selected cDNA, 3 μl USER Enzyme (NEB, MA, USA) was added and heated to 37 °C for 15 min followed by 5 min at 95 °C.

    Plasmid Preparation:

    Article Title: Who Needs Neighbors? PKS8 Is a Stand-Alone Gene in Fusarium graminearum Responsible for Production of Gibepyrones and Prolipyrone B
    Article Snippet: .. The PCR fragments were cloned into a linearized pRF-HU2E vector under control of the gpdA promoter by a four fragment cloning step using the USER enzyme™ (New England Biolabs, Ipswich, MA, USA) and verified by colony PCR [ ]. .. Transformation of F. graminearum was carried out by Agrobacterium tumefaciens mediated transformation as described previously [ ] and the resulting mutants were verified by diagnostic PCR using a forward primer annealing to gDNA outside the border region and a reverse primer annealing to the hygromycin resistance gene.

    Article Title: Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
    Article Snippet: .. PCR products were purified with Illustra GFX PCR DNA and a Gel Band Purification Kit (GE Healthcare, UK) and ligated to linearized vector pRF-HU2 with USER enzyme (New England BioLabs, USA). .. Vector constructs were multiplied in E . coli DH5α cells and isolated with a High Pure Plasmid Isolation Kit (Roche, Life Science, USA).

    Article Title: Continuous directed evolution of aminoacyl-tRNA synthetases
    Article Snippet: Plasmids and selection phage were prepared using isothermal assembly with Gibson Assembly 2x Master Mix (New England Biolabs), USER cloning with USER enzyme (New England Biolabs), or ligation cycling reaction with Ampligase (Epicentre). .. Chimeric PylRS, Mb PylRS, and Mm PylRS were obtained from pTECH plasmid sources from previous studies.

    RNA Extraction:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: Paragraph title: 2.2. RNA Extraction, Library Preparation, Sequencing and De Novo Assembly ... The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C.

    Article Title: Transcriptome and gene expression analysis of DHA producer Aurantiochytrium under low temperature conditions
    Article Snippet: Paragraph title: RNA extraction, library preparation and sequencing ... Then, 3 μL of USER enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37 °C for 15 min, followed by 5 min at 95 °C before PCR.

    Selection:

    Article Title: Continuous directed evolution of aminoacyl-tRNA synthetases
    Article Snippet: .. Plasmids and selection phage were prepared using isothermal assembly with Gibson Assembly 2x Master Mix (New England Biolabs), USER cloning with USER enzyme (New England Biolabs), or ligation cycling reaction with Ampligase (Epicentre). .. Genes were either synthesized from gBlock gene fragments (Integrated DNA Technologies) or PCR amplified from native sources.

    Sample Prep:

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: Second-strand cDNA was synthesised by adding 91.8 μl H2 O, 5 μg random hexamers, 4 μl 5× first-strand buffer, 2 μl 100 mM DTT, 4 μl 10 mM dNTPs with dTTP replaced by dUTP, 30 μl 5× second-strand buffer, 40 U E. coli DNA polymerase, 10 U E . coli DNA ligase and 2 U E . coli RNase H, and incubated at 16 °C for 2 h, followed by incubation with 10 U T4 polymerase at 16 °C for 10 min. Double-stranded cDNA was purified using a Qiagen MinElute column and used for Illumina sample preparation and sequencing according to the Illumina protocol. .. Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C.

    Ethanol Precipitation:

    Article Title: Integrative Analyses of Long Non-coding RNA and mRNA Involved in Piglet Ileum Immune Response to Clostridium perfringens Type C Infection
    Article Snippet: Ribosomal RNA (rRNA) was removed from total RNA by epicenter Ribo-zero™ rRNA Removal Kit (epicenter, USA), and rRNA free residue was cleaned up by ethanol precipitation. .. Then 3 μL USER Enzyme (NEB, USA) was used with size-selected, adaptor-ligated cDNA at 37°C for 15 min followed by 5 min at 95°C before PCR, then PCR reaction was performed using Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer.

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: Sixteen microlitres of purified RNA was fragmented by addition of 4 μl 5× fragmentation buffer (200 mM Tris acetate (pH 8.2), 500 mM potassium acetate and 150 mM magnesium acetate) and incubated at 94 °C for exactly 90 s. After ethanol precipitation, fragment ed RNA was mixed with 5 μg random hexamers, followed by incubation at 70 °C for 10 min and chilling on ice. .. Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C.

    Incubation:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: .. The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C. .. Then PCR reaction was performed using Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer.

    Article Title: Screening and characterization of long noncoding RNAs involved in the albinism of Ananas comosus var. bracteatus leaves
    Article Snippet: .. Then, 3 μl of USER enzyme (NEB, USA) was incubated with the size-selected, adaptor-ligated cDNA at 37°C for 15 min before PCR. .. PCR was then performed with Phusion high-fidelity DNA polymerase, universal PCR primers and the index (X) primer.

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: .. Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C. .. The 300-bp libraries were used for cluster generation on a HiSeq 2000 (Illumina).

    Article Title: Mitogenome evolution in the last surviving woolly mammoth population reveals neutral and functional consequences of small population size
    Article Snippet: Double stranded Illumina libraries were prepared from 20 μL of DNA extract according to Meyer and Kircher , using uracil‐treatment with the USER enzyme (New England Biolabs; Briggs et al. ). .. T4 DNA polymerase was added to the reaction mix following a three‐hour incubation at 37°C.

    Knock-Out:

    Article Title: Comprehensive analysis of Verticillium nonalfalfae in silico secretome uncovers putative effector proteins expressed during hop invasion
    Article Snippet: Paragraph title: Construction of knockout vectors and preparation of V . nonalfalfae knockout mutants ... PCR products were purified with Illustra GFX PCR DNA and a Gel Band Purification Kit (GE Healthcare, UK) and ligated to linearized vector pRF-HU2 with USER enzyme (New England BioLabs, USA).

    Produced:

    Article Title: De Novo Transcriptome Assembly and Population Genetic Analyses for an Endangered Chinese Endemic Acer miaotaiense (Aceraceae)
    Article Snippet: First, strand cDNA was produced based on the random hexamer primer and M-MuLV Reverse Transcriptase. .. The fragments in library were purified with AMPure XP system (Beckman Coulter, Danvers, MA, USA) to firstly select for 240 bp cDNA fragments; 3 μL USER Enzyme (New England Biolabs) was incubated with size-selected, adaptor-ligated cDNA for 15 min at 37 °C and then 5 min at 95 °C.

    Concentration Assay:

    Article Title: E-cadherin loss induces targetable autocrine activation of growth factor signalling in lobular breast cancer
    Article Snippet: After measurement of RNA concentration using a Qubit fluorometer (Invitrogen), 250 ng of total RNA was treated using a Ribo-Zero rRNA removal kit (Epicenter) to remove ribosomal RNAs. .. Before the final PCR, a band corresponding to ~300 bp (DNA + adaptor) was collected and incubated with 1 U USER enzyme (NEB) at 37 °C for 15 min, followed by 5 min at 95 °C.

    Article Title: Mitogenome evolution in the last surviving woolly mammoth population reveals neutral and functional consequences of small population size
    Article Snippet: Double stranded Illumina libraries were prepared from 20 μL of DNA extract according to Meyer and Kircher , using uracil‐treatment with the USER enzyme (New England Biolabs; Briggs et al. ). .. During the blunt‐end repair, USER enzyme was added so that the final concentration was 0.15 U/μL in the reaction mix described in “Step 4” of Meyer and Kircher ( ).

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    New England Biolabs user enzyme
    User Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 403 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/user enzyme/product/New England Biolabs
    Average 90 stars, based on 403 article reviews
    Price from $9.99 to $1999.99
    user enzyme - by Bioz Stars, 2020-01
    90/100 stars
      Buy from Supplier

    Image Search Results