r3140  (New England Biolabs)


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  • 90
    Name:
    PstI HF
    Description:
    PstI HF 50 000 units
    Catalog Number:
    r3140l
    Price:
    264
    Size:
    50 000 units
    Category:
    Restriction Enzymes
    Buy from Supplier


    Structured Review

    New England Biolabs r3140
    PstI HF
    PstI HF 50 000 units
    https://www.bioz.com/result/r3140/product/New England Biolabs
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    r3140 - by Bioz Stars, 2020-01
    90/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
    Article Snippet: All the standard parts were domesticated for Aar I and Bsa I when necessary and were cloned into mUAV. .. Subsequently the pSB1C3/pSB1K3 backbones and the Golden Gate cassettes were digested with EcoRI-HF and PstI-HF (NEB) for 20 min at 37°C followed by purification with PCR Clean-up kit (Macherey Nagel).

    Amplification:

    Article Title: Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
    Article Snippet: They were then amplified using Q5 DNA polymerase (NEB) with primers bearing EcoRI , AarI , BsaI , and PstI restriction sites and 4 bp overhangs. .. Subsequently the pSB1C3/pSB1K3 backbones and the Golden Gate cassettes were digested with EcoRI-HF and PstI-HF (NEB) for 20 min at 37°C followed by purification with PCR Clean-up kit (Macherey Nagel).

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: G-blocks were PCR amplified using Forward primer 5′-TTATTCGAATTCGCGGCCGCTTCTAGAG and Reverse primer 5′-GGATTTCTGCAGCGGCCGCTACTAGTA with the following conditions: in a 50 μL reaction: 10 μL 5x HF buffer (New England Biolabs, NEB #E0553L), 2 μL 10 mM dNTP mix (NEB #N0447L), 1 μL 100 μM forward and reverse primer, 25 ng g block template, and 0.5 μL phusion polymerase. .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT).

    Synthesized:

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: Gene cassettes were designed in CLC Main (Qiagen) and synthesized as g-blocks by IDT. .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT).

    Blocking Assay:

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: G-blocks were PCR amplified using Forward primer 5′-TTATTCGAATTCGCGGCCGCTTCTAGAG and Reverse primer 5′-GGATTTCTGCAGCGGCCGCTACTAGTA with the following conditions: in a 50 μL reaction: 10 μL 5x HF buffer (New England Biolabs, NEB #E0553L), 2 μL 10 mM dNTP mix (NEB #N0447L), 1 μL 100 μM forward and reverse primer, 25 ng g block template, and 0.5 μL phusion polymerase. .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT).

    Electrophoresis:

    Article Title: Construction of a high-density linkage map and fine mapping of QTL for growth in Asian seabass
    Article Snippet: 200 ng of DNA from each sample was digested with 20 units of PstI-HF and MspI restriction enzymes (New England Biolabs, USA). .. Digested DNA fragments were examined by electrophoresis and were then ligated with barcoded adaptors .

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging
    Article Snippet: .. DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe. .. The DNA was transferred to a NYTRAN-N membrane (GE Healthcare) by capillary action in 20× SSC (1× SSC is 0.15 M NaCl plus 0.015 M sodium citrate) overnight and cross-linked to the membrane in a StrataLinker 2400 (Stratagene) using the AutoUV setting.

    Article Title: Fine mapping QTL for resistance to VNN disease using a high-density linkage map in Asian seabass
    Article Snippet: In brief, 200 ng of each DNA was digested with 20 units of restriction enzyme of PstI-HF and MspI (New England Biolabs, Ipswich, MA, USA) at 37 °C for 2.5 hours. .. The DNA fragments were examined by electrophoresis on a 2% agarose gel before ligation with barcoded adaptors .

    Incubation:

    Article Title: Single-molecule imaging reveals the mechanism of Exo1 regulation by single-stranded DNA binding proteins
    Article Snippet: .. Resection assays were conducted as above, but with a few modifications. hExo1-bio or yExo1-bio at indicated concentrations were incubated in imaging buffer with 10 ng PstI-HF (4-nt 3′ overhang; NEB) linearized 6.3-kb DNA in the presence or absence of hRPA or yRPA as indicated for 1 h at 37 °C or 30 °C. .. The reactions were deproteinized with 2 µg Proteinase K (NEB), run on a gel, stained, and analyzed as above.

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging
    Article Snippet: BAC16.iSLK cells were harvested in proteinase K digestion buffer and incubated overnight with proteinase K (80 μg/ml). .. DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe.

    In Silico:

    Article Title: Whole Genome Amplification and Reduced-Representation Genome Sequencing of Schistosoma japonicum Miracidia
    Article Snippet: .. Correspondence between in silico and empirical results Based on comparisons of in silico digestions of the complete genome sequence of S . japonicum [ ] using different potential pairs of restriction enzymes, we chose the combination of PstI -HF and Sau3AI for empirical ddRADseq library construction and sequencing. ..

    Modification:

    Article Title: Chromosome‐level assembly, genetic and physical mapping of Phalaenopsis aphrodite genome provides new insights into species adaptation and resources for orchid breeding
    Article Snippet: One microgram gDNA was digested with 20 units of PstI ‐HF (New England BioLabs [NEB], Ipswish, MA) overnight in a 50 μL reaction volume. .. Digested DNA was ligated to 2 μL 100 nm P1 barcoded adapter, which is a modified Solexa adapter, along with 1 μL 10× NEBuffer4 (NEB), 0.5 μL 2000 unit/μL T4 DNA ligase (NEB), and 0.6 μL 100 mm riboATP (Promega, Madison, WI) in a 60 μL reaction volume for 1 h. Samples were heat‐inactivated for 20 min at 65 °C.

    Article Title: Reassessment of QTLs for Late Blight Resistance in the Tomato Accession L3708 Using a Restriction Site Associated DNA (RAD) Linkage Map and Highly Aggressive Isolates of Phytophthora infestans
    Article Snippet: Construction of the RAD library Total genomic DNA was isolated from young tomato leaves using a modified CTAB method , and further purified using the DNeasy Blood & Tissue Kit (QIAGEN, Venlo, Netherland) following manufacturer's instructions. .. For each sample, 1 µg gDNA was digested with 20 units of PstI -HF (New England BioLabs [NEB], Ipswish MA, USA) overnight in a 50 µL reaction volume.

    Article Title: Chromosome‐level assembly, genetic and physical mapping of Phalaenopsis aphrodite genome provides new insights into species adaptation and resources for orchid breeding
    Article Snippet: One microgram gDNA was digested with 20 units of PstI ‐HF (New England BioLabs [NEB], Ipswish, MA) overnight in a 50 μL reaction volume. .. Digested DNA was ligated to 2 μL 100 n m P1 barcoded adapter, which is a modified Solexa adapter, along with 1 μL 10× NEBuffer4 (NEB), 0.5 μL 2000 unit/μL T4 DNA ligase (NEB), and 0.6 μL 100 m m riboATP (Promega, Madison, WI) in a 60 μL reaction volume for 1 h. Samples were heat‐inactivated for 20 min at 65 °C.

    Transformation Assay:

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT). .. The ligation was transformed into chemically competent BL21 E. coli (NEB #C2530H) with the following conditions: 5 μL of ligation into 50 μL cells on ice for 5 min, followed by 45 s at 42°C, followed by 2 min on ice.

    Recombinase Polymerase Amplification:

    Article Title: Single-molecule imaging reveals the mechanism of Exo1 regulation by single-stranded DNA binding proteins
    Article Snippet: Paragraph title: Gel-based resection assays with Exo1 and RPA. ... Resection assays were conducted as above, but with a few modifications. hExo1-bio or yExo1-bio at indicated concentrations were incubated in imaging buffer with 10 ng PstI-HF (4-nt 3′ overhang; NEB) linearized 6.3-kb DNA in the presence or absence of hRPA or yRPA as indicated for 1 h at 37 °C or 30 °C.

    Derivative Assay:

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging
    Article Snippet: DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe. .. Briefly, the membrane was hybridized at 42°C in EasyHyb buffer with a DIG-labeled linear terminal repeat subunit derived from AscI (New England BioLabs) digested pK8TR, which was labeled according to kit instructions.

    Southern Blot:

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging
    Article Snippet: Paragraph title: Southern blotting. ... DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe.

    Ligation:

    Article Title: Construction of a high-density linkage map and fine mapping of QTL for growth in Asian seabass
    Article Snippet: 200 ng of DNA from each sample was digested with 20 units of PstI-HF and MspI restriction enzymes (New England Biolabs, USA). .. Ligation products were pooled for size selection of 300–500 bp using Pippin Prep (Sage Science, USA) after clean up with QIAquick PCR Purification Kit (Qiagen, Germany).

    Article Title: Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
    Article Snippet: Subsequently the pSB1C3/pSB1K3 backbones and the Golden Gate cassettes were digested with EcoRI-HF and PstI-HF (NEB) for 20 min at 37°C followed by purification with PCR Clean-up kit (Macherey Nagel). .. Ligation was mediated by T7 DNA ligase (NEB) to construct mUAV, Level 1 and Level 2 Acceptor Vectors.

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT). .. The ligation was transformed into chemically competent BL21 E. coli (NEB #C2530H) with the following conditions: 5 μL of ligation into 50 μL cells on ice for 5 min, followed by 45 s at 42°C, followed by 2 min on ice.

    Article Title: Fine mapping QTL for resistance to VNN disease using a high-density linkage map in Asian seabass
    Article Snippet: In brief, 200 ng of each DNA was digested with 20 units of restriction enzyme of PstI-HF and MspI (New England Biolabs, Ipswich, MA, USA) at 37 °C for 2.5 hours. .. The DNA fragments were examined by electrophoresis on a 2% agarose gel before ligation with barcoded adaptors .

    Imaging:

    Article Title: Single-molecule imaging reveals the mechanism of Exo1 regulation by single-stranded DNA binding proteins
    Article Snippet: .. Resection assays were conducted as above, but with a few modifications. hExo1-bio or yExo1-bio at indicated concentrations were incubated in imaging buffer with 10 ng PstI-HF (4-nt 3′ overhang; NEB) linearized 6.3-kb DNA in the presence or absence of hRPA or yRPA as indicated for 1 h at 37 °C or 30 °C. .. The reactions were deproteinized with 2 µg Proteinase K (NEB), run on a gel, stained, and analyzed as above.

    DNA Labeling:

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging
    Article Snippet: DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe. .. The membrane was treated according to the DIG High Prime DNA labeling and detection starter kit II (Roche) according to the manufacturer's instructions.

    Sequencing:

    Article Title: Construction of a high-density linkage map and fine mapping of QTL for growth in Asian seabass
    Article Snippet: Paragraph title: RAD library preparation and sequencing ... 200 ng of DNA from each sample was digested with 20 units of PstI-HF and MspI restriction enzymes (New England Biolabs, USA).

    Article Title: Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
    Article Snippet: Subsequently the pSB1C3/pSB1K3 backbones and the Golden Gate cassettes were digested with EcoRI-HF and PstI-HF (NEB) for 20 min at 37°C followed by purification with PCR Clean-up kit (Macherey Nagel). .. To construct the 50 bp linkers in the Auxiliary Plasmids, a short sequence was PCR amplified from scOrange gene using primers that contained the appropriate overhangs and the Aar I and Bsa I recogni tion sites.

    Article Title: Genomic diversity in switchgrass (Panicum virgatum): from the continental scale to a dune landscape
    Article Snippet: .. To produce the second set of RRLs (Sequencing run 2, ), genomic DNA (5.0 ug per sample) was digested with PstI-HF (NEB, Ipswich MA), then processed according to the Illumina protocol for preparing libraries for multiplexed paired-end sequencing without the DNA nebulization step. .. These libraries were sequenced with 100bp paired-end runs on an Illumina GA-II.

    Article Title: Whole Genome Amplification and Reduced-Representation Genome Sequencing of Schistosoma japonicum Miracidia
    Article Snippet: .. Correspondence between in silico and empirical results Based on comparisons of in silico digestions of the complete genome sequence of S . japonicum [ ] using different potential pairs of restriction enzymes, we chose the combination of PstI -HF and Sau3AI for empirical ddRADseq library construction and sequencing. ..

    Article Title: Fine mapping QTL for resistance to VNN disease using a high-density linkage map in Asian seabass
    Article Snippet: Paragraph title: Sequencing library preparation and next generation sequencing ... In brief, 200 ng of each DNA was digested with 20 units of restriction enzyme of PstI-HF and MspI (New England Biolabs, Ipswich, MA, USA) at 37 °C for 2.5 hours.

    Nucleic Acid Electrophoresis:

    Article Title: Evolved Populations of Shigella flexneri Phage Sf6 Acquire Large Deletions, Altered Genomic Architecture, and Faster Life Cycles
    Article Snippet: .. Restriction Enzyme Digests to Show Terminal Redundancy Single restriction enzyme digests used BamHI HF and PstI HF (New England Biolabs) at 37°C for 15 min. Gel electrophoresis used 0.7% agarose in TAE and was run for 90 min at a constant 80 V. The ladder used was 1 kb Plus (Invitrogen). ..

    Isolation:

    Article Title: Chromosome‐level assembly, genetic and physical mapping of Phalaenopsis aphrodite genome provides new insights into species adaptation and resources for orchid breeding
    Article Snippet: Total genomic DNA was isolated from young flower buds using the Plant Mini Kit (Qiagen, Venlo, The Netherlands) following the manufacturer's instructions. .. One microgram gDNA was digested with 20 units of PstI ‐HF (New England BioLabs [NEB], Ipswish, MA) overnight in a 50 μL reaction volume.

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging
    Article Snippet: DNA was isolated by phenol-chloroform extraction and ethanol precipitation and then resuspended in TE buffer. .. DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe.

    Article Title: Reassessment of QTLs for Late Blight Resistance in the Tomato Accession L3708 Using a Restriction Site Associated DNA (RAD) Linkage Map and Highly Aggressive Isolates of Phytophthora infestans
    Article Snippet: Construction of the RAD library Total genomic DNA was isolated from young tomato leaves using a modified CTAB method , and further purified using the DNeasy Blood & Tissue Kit (QIAGEN, Venlo, Netherland) following manufacturer's instructions. .. For each sample, 1 µg gDNA was digested with 20 units of PstI -HF (New England BioLabs [NEB], Ipswish MA, USA) overnight in a 50 µL reaction volume.

    Article Title: Chromosome‐level assembly, genetic and physical mapping of Phalaenopsis aphrodite genome provides new insights into species adaptation and resources for orchid breeding
    Article Snippet: Total genomic DNA was isolated from young flower buds using the Plant Mini Kit (Qiagen, Venlo, The Netherlands) following the manufacturer's instructions. .. One microgram gDNA was digested with 20 units of PstI ‐HF (New England BioLabs [NEB], Ipswish, MA) overnight in a 50 μL reaction volume.

    Labeling:

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging
    Article Snippet: DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe. .. Briefly, the membrane was hybridized at 42°C in EasyHyb buffer with a DIG-labeled linear terminal repeat subunit derived from AscI (New England BioLabs) digested pK8TR, which was labeled according to kit instructions.

    Purification:

    Article Title: Construction of a high-density linkage map and fine mapping of QTL for growth in Asian seabass
    Article Snippet: 200 ng of DNA from each sample was digested with 20 units of PstI-HF and MspI restriction enzymes (New England Biolabs, USA). .. Ligation products were pooled for size selection of 300–500 bp using Pippin Prep (Sage Science, USA) after clean up with QIAquick PCR Purification Kit (Qiagen, Germany).

    Article Title: Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
    Article Snippet: .. Subsequently the pSB1C3/pSB1K3 backbones and the Golden Gate cassettes were digested with EcoRI-HF and PstI-HF (NEB) for 20 min at 37°C followed by purification with PCR Clean-up kit (Macherey Nagel). .. Ligation was mediated by T7 DNA ligase (NEB) to construct mUAV, Level 1 and Level 2 Acceptor Vectors.

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: PCR products were column purified (Qiagen PCR cleanup kit #28104), and 2 μg of DNA was digested with EcoRI-HF and PstI-HF (NEB) with the manufacturer's recommended conditions, followed by another PCR cleanup (Qiagen #28104). .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT).

    Article Title: Whole Genome Amplification and Reduced-Representation Genome Sequencing of Schistosoma japonicum Miracidia
    Article Snippet: Library preparation and quality assessment Adult worm DNA or whole-genome-amplified miracidium DNA was digested with two restriction enzymes, PstI -HF (New England Biolabs (NEB) R3140), a 6-cutter, and Sau3AI (NEB R0169), a 4-cutter, for eight hours at 37°C. .. Following digestions, DNA was purified via solid phase reversible immobilization (SPRI) using Axygen AxyPrep paramagnetic beads.

    Article Title: Fine mapping QTL for resistance to VNN disease using a high-density linkage map in Asian seabass
    Article Snippet: In brief, 200 ng of each DNA was digested with 20 units of restriction enzyme of PstI-HF and MspI (New England Biolabs, Ipswich, MA, USA) at 37 °C for 2.5 hours. .. The ligation products were pooled, followed by the size selection of 350 to 600 bp using Pippin Prep (Sage Science, Beverly, MA, USA), and then a cleanup using QIAquick PCR Purification Kit (Qiagen, Hilden, Germany).

    Article Title: Reassessment of QTLs for Late Blight Resistance in the Tomato Accession L3708 Using a Restriction Site Associated DNA (RAD) Linkage Map and Highly Aggressive Isolates of Phytophthora infestans
    Article Snippet: Construction of the RAD library Total genomic DNA was isolated from young tomato leaves using a modified CTAB method , and further purified using the DNeasy Blood & Tissue Kit (QIAGEN, Venlo, Netherland) following manufacturer's instructions. .. For each sample, 1 µg gDNA was digested with 20 units of PstI -HF (New England BioLabs [NEB], Ipswish MA, USA) overnight in a 50 µL reaction volume.

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: .. PCR products were column purified (Qiagen PCR cleanup kit #28104), and 2 μg of DNA was digested with EcoRI-HF and PstI-HF (NEB) with the manufacturer's recommended conditions, followed by another PCR cleanup (Qiagen #28104). .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT).

    Polymerase Chain Reaction:

    Article Title: Construction of a high-density linkage map and fine mapping of QTL for growth in Asian seabass
    Article Snippet: 200 ng of DNA from each sample was digested with 20 units of PstI-HF and MspI restriction enzymes (New England Biolabs, USA). .. Ligation products were pooled for size selection of 300–500 bp using Pippin Prep (Sage Science, USA) after clean up with QIAquick PCR Purification Kit (Qiagen, Germany).

    Article Title: Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
    Article Snippet: .. Subsequently the pSB1C3/pSB1K3 backbones and the Golden Gate cassettes were digested with EcoRI-HF and PstI-HF (NEB) for 20 min at 37°C followed by purification with PCR Clean-up kit (Macherey Nagel). .. Ligation was mediated by T7 DNA ligase (NEB) to construct mUAV, Level 1 and Level 2 Acceptor Vectors.

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: PCR products were column purified (Qiagen PCR cleanup kit #28104), and 2 μg of DNA was digested with EcoRI-HF and PstI-HF (NEB) with the manufacturer's recommended conditions, followed by another PCR cleanup (Qiagen #28104). .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT).

    Article Title: Chromosome‐level assembly, genetic and physical mapping of Phalaenopsis aphrodite genome provides new insights into species adaptation and resources for orchid breeding
    Article Snippet: One microgram gDNA was digested with 20 units of PstI ‐HF (New England BioLabs [NEB], Ipswish, MA) overnight in a 50 μL reaction volume. .. Each pooled sample was divided into several 50 μL aliquots in 0.5‐mL PCR tubes (Axygen catalog # PCR‐05‐C, Corning, Tewksbury, MA).

    Article Title: Fine mapping QTL for resistance to VNN disease using a high-density linkage map in Asian seabass
    Article Snippet: In brief, 200 ng of each DNA was digested with 20 units of restriction enzyme of PstI-HF and MspI (New England Biolabs, Ipswich, MA, USA) at 37 °C for 2.5 hours. .. The ligation products were pooled, followed by the size selection of 350 to 600 bp using Pippin Prep (Sage Science, Beverly, MA, USA), and then a cleanup using QIAquick PCR Purification Kit (Qiagen, Hilden, Germany).

    Article Title: Reassessment of QTLs for Late Blight Resistance in the Tomato Accession L3708 Using a Restriction Site Associated DNA (RAD) Linkage Map and Highly Aggressive Isolates of Phytophthora infestans
    Article Snippet: For each sample, 1 µg gDNA was digested with 20 units of PstI -HF (New England BioLabs [NEB], Ipswish MA, USA) overnight in a 50 µL reaction volume. .. Samples were pooled (20 µL each, 60 samples) and a 50 µL aliquot was loaded into a 0.5 mL PCR tube (Axygen catalog # PCR-05-C, Corning, Tewksbury, MA, USA).

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: .. PCR products were column purified (Qiagen PCR cleanup kit #28104), and 2 μg of DNA was digested with EcoRI-HF and PstI-HF (NEB) with the manufacturer's recommended conditions, followed by another PCR cleanup (Qiagen #28104). .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT).

    Article Title: Chromosome‐level assembly, genetic and physical mapping of Phalaenopsis aphrodite genome provides new insights into species adaptation and resources for orchid breeding
    Article Snippet: One microgram gDNA was digested with 20 units of PstI ‐HF (New England BioLabs [NEB], Ipswish, MA) overnight in a 50 μL reaction volume. .. Each pooled sample was divided into several 50 μL aliquots in 0.5‐mL PCR tubes (Axygen catalog # PCR‐05‐C, Corning, Tewksbury, MA).

    Construct:

    Article Title: Construction of a high-density linkage map and fine mapping of QTL for growth in Asian seabass
    Article Snippet: RAD libraries were constructed using the double digest RADseq method with some modifications . .. 200 ng of DNA from each sample was digested with 20 units of PstI-HF and MspI restriction enzymes (New England Biolabs, USA).

    Article Title: Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
    Article Snippet: Subsequently the pSB1C3/pSB1K3 backbones and the Golden Gate cassettes were digested with EcoRI-HF and PstI-HF (NEB) for 20 min at 37°C followed by purification with PCR Clean-up kit (Macherey Nagel). .. Ligation was mediated by T7 DNA ligase (NEB) to construct mUAV, Level 1 and Level 2 Acceptor Vectors.

    Plasmid Preparation:

    Article Title: Mobius Assembly: A versatile Golden-Gate framework towards universal DNA assembly
    Article Snippet: Paragraph title: Part and vector generation ... Subsequently the pSB1C3/pSB1K3 backbones and the Golden Gate cassettes were digested with EcoRI-HF and PstI-HF (NEB) for 20 min at 37°C followed by purification with PCR Clean-up kit (Macherey Nagel).

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT). .. Purified, digested insert and vector were ligated with the following conditions 1 μL 10X T4 DNA ligase buffer (NEB), 50 ng vector, 150 ng insert, 0.5 μL T4 DNA ligase (NEB) in 10 μL total for 10 min at room temperature.

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: Paragraph title: Construction of Plasmid DNA ... PCR products were column purified (Qiagen PCR cleanup kit #28104), and 2 μg of DNA was digested with EcoRI-HF and PstI-HF (NEB) with the manufacturer's recommended conditions, followed by another PCR cleanup (Qiagen #28104).

    Selection:

    Article Title: Construction of a high-density linkage map and fine mapping of QTL for growth in Asian seabass
    Article Snippet: 200 ng of DNA from each sample was digested with 20 units of PstI-HF and MspI restriction enzymes (New England Biolabs, USA). .. Ligation products were pooled for size selection of 300–500 bp using Pippin Prep (Sage Science, USA) after clean up with QIAquick PCR Purification Kit (Qiagen, Germany).

    Article Title: Fine mapping QTL for resistance to VNN disease using a high-density linkage map in Asian seabass
    Article Snippet: In brief, 200 ng of each DNA was digested with 20 units of restriction enzyme of PstI-HF and MspI (New England Biolabs, Ipswich, MA, USA) at 37 °C for 2.5 hours. .. The ligation products were pooled, followed by the size selection of 350 to 600 bp using Pippin Prep (Sage Science, Beverly, MA, USA), and then a cleanup using QIAquick PCR Purification Kit (Qiagen, Hilden, Germany).

    Agarose Gel Electrophoresis:

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT). .. Purified, digested insert and vector were ligated with the following conditions 1 μL 10X T4 DNA ligase buffer (NEB), 50 ng vector, 150 ng insert, 0.5 μL T4 DNA ligase (NEB) in 10 μL total for 10 min at room temperature.

    Article Title: Fine mapping QTL for resistance to VNN disease using a high-density linkage map in Asian seabass
    Article Snippet: In brief, 200 ng of each DNA was digested with 20 units of restriction enzyme of PstI-HF and MspI (New England Biolabs, Ipswich, MA, USA) at 37 °C for 2.5 hours. .. The DNA fragments were examined by electrophoresis on a 2% agarose gel before ligation with barcoded adaptors .

    Article Title: Engineered Orthogonal Quorum Sensing Systems for Synthetic Gene Regulation in Escherichia coli
    Article Snippet: PCR products were column purified (Qiagen PCR cleanup kit #28104), and 2 μg of DNA was digested with EcoRI-HF and PstI-HF (NEB) with the manufacturer's recommended conditions, followed by another PCR cleanup (Qiagen #28104). .. Six micrograms of backbone vector pSB1A3 was digested with EcoRI-HF and PstI-HF (NEB), electrophoresed on a 1% agarose gel for 45 min, and gel extracted (Sigma #NA1111-1KT).

    Ethanol Precipitation:

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging
    Article Snippet: DNA was isolated by phenol-chloroform extraction and ethanol precipitation and then resuspended in TE buffer. .. DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe.

    Next-Generation Sequencing:

    Article Title: Fine mapping QTL for resistance to VNN disease using a high-density linkage map in Asian seabass
    Article Snippet: Paragraph title: Sequencing library preparation and next generation sequencing ... In brief, 200 ng of each DNA was digested with 20 units of restriction enzyme of PstI-HF and MspI (New England Biolabs, Ipswich, MA, USA) at 37 °C for 2.5 hours.

    Concentration Assay:

    Article Title: Construction of a high-density linkage map and fine mapping of QTL for growth in Asian seabass
    Article Snippet: RAD library preparation and sequencing Genomic DNA concentration was measured using Qubit® assays (Life Technologies, USA). .. 200 ng of DNA from each sample was digested with 20 units of PstI-HF and MspI restriction enzymes (New England Biolabs, USA).

    High Throughput Screening Assay:

    Article Title: Genomic diversity in switchgrass (Panicum virgatum): from the continental scale to a dune landscape
    Article Snippet: Paragraph title: Library preparation and high-throughput sequencing ... To produce the second set of RRLs (Sequencing run 2, ), genomic DNA (5.0 ug per sample) was digested with PstI-HF (NEB, Ipswich MA), then processed according to the Illumina protocol for preparing libraries for multiplexed paired-end sequencing without the DNA nebulization step.

    Staining:

    Article Title: Single-molecule imaging reveals the mechanism of Exo1 regulation by single-stranded DNA binding proteins
    Article Snippet: Resection assays were conducted as above, but with a few modifications. hExo1-bio or yExo1-bio at indicated concentrations were incubated in imaging buffer with 10 ng PstI-HF (4-nt 3′ overhang; NEB) linearized 6.3-kb DNA in the presence or absence of hRPA or yRPA as indicated for 1 h at 37 °C or 30 °C. .. The reactions were deproteinized with 2 µg Proteinase K (NEB), run on a gel, stained, and analyzed as above.

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging
    Article Snippet: .. DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe. .. The DNA was transferred to a NYTRAN-N membrane (GE Healthcare) by capillary action in 20× SSC (1× SSC is 0.15 M NaCl plus 0.015 M sodium citrate) overnight and cross-linked to the membrane in a StrataLinker 2400 (Stratagene) using the AutoUV setting.

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    New England Biolabs r3140
    ORF68 PTC virus displays a genome cleavage defect. (A) Diagram depicting the <t>DNA</t> cleavage assay protocol and the expected TR DNA laddering phenotype (or lack thereof) upon infection with WT or ORF68 PTC virus. (B) Southern blot of the <t>PstI-HF</t> digested DNA with a TR probe from cells infected with WT, ORF68 PTC , or ORF68 PTC -MR virus.
    R3140, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ORF68 PTC virus displays a genome cleavage defect. (A) Diagram depicting the DNA cleavage assay protocol and the expected TR DNA laddering phenotype (or lack thereof) upon infection with WT or ORF68 PTC virus. (B) Southern blot of the PstI-HF digested DNA with a TR probe from cells infected with WT, ORF68 PTC , or ORF68 PTC -MR virus.

    Journal: Journal of Virology

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus ORF68 Is a DNA Binding Protein Required for Viral Genome Cleavage and Packaging

    doi: 10.1128/JVI.00840-18

    Figure Lengend Snippet: ORF68 PTC virus displays a genome cleavage defect. (A) Diagram depicting the DNA cleavage assay protocol and the expected TR DNA laddering phenotype (or lack thereof) upon infection with WT or ORF68 PTC virus. (B) Southern blot of the PstI-HF digested DNA with a TR probe from cells infected with WT, ORF68 PTC , or ORF68 PTC -MR virus.

    Article Snippet: DNA (10 μg) was digested with PstI-HF (New England BioLabs) overnight then separated by electrophoresis in a 0.7% agarose 1× TBE gel stained with SYBRsafe.

    Techniques: DNA Cleavage Assay, DNA Laddering, Infection, Southern Blot