nhei  (New England Biolabs)


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    Structured Review

    New England Biolabs nhei
    Nhei, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nhei/product/New England Biolabs
    Average 99 stars, based on 27 article reviews
    Price from $9.99 to $1999.99
    nhei - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Article Title: A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If)
    Article Snippet: .. The cDNA generated by the PCR reaction described above using SL35-F-(13F) and SL35-R-(13R) was digested with NheI (New England BioLabs Inc.) and cloned into the expression vector pTre2hyg (CLONTECH Laboratories Inc., Palo Alto, California, USA) that had been digested with NheI and EcoRV (New England BioLabs Inc.). .. After amplification in bacteria (TOP10F′; Invitrogen Corp., San Diego, California, USA), the vector constructs were purified, and the sequence of the coding region was verified by direct sequencing using a LI-COR sequencer (LI-COR Biosciences, Bad Homburg, Germany).

    Amplification:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Article Title: A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If)
    Article Snippet: The cDNA generated by the PCR reaction described above using SL35-F-(13F) and SL35-R-(13R) was digested with NheI (New England BioLabs Inc.) and cloned into the expression vector pTre2hyg (CLONTECH Laboratories Inc., Palo Alto, California, USA) that had been digested with NheI and EcoRV (New England BioLabs Inc.). .. After amplification in bacteria (TOP10F′; Invitrogen Corp., San Diego, California, USA), the vector constructs were purified, and the sequence of the coding region was verified by direct sequencing using a LI-COR sequencer (LI-COR Biosciences, Bad Homburg, Germany).

    Polymerase Chain Reaction:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Article Title: A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If)
    Article Snippet: .. The cDNA generated by the PCR reaction described above using SL35-F-(13F) and SL35-R-(13R) was digested with NheI (New England BioLabs Inc.) and cloned into the expression vector pTre2hyg (CLONTECH Laboratories Inc., Palo Alto, California, USA) that had been digested with NheI and EcoRV (New England BioLabs Inc.). .. After amplification in bacteria (TOP10F′; Invitrogen Corp., San Diego, California, USA), the vector constructs were purified, and the sequence of the coding region was verified by direct sequencing using a LI-COR sequencer (LI-COR Biosciences, Bad Homburg, Germany).

    Mutagenesis:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: The vpu mutant clone pMJ4-Vpu-S58,62N was constructed using QuikChange. .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB).

    Introduce:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Purification:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Article Title: Directed evolution of a picomolar-affinity, high-specificity antibody targeting phosphorylated tau
    Article Snippet: Paragraph title: Purification of pThr-231 scFvs ... The pThr-231 scFv gene was inserted into the vector pRS316 (a gift from Dr. Eric Shusta), which had been restriction-digested with NheI and BsrGI (New England Biolabs).

    Article Title: A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If)
    Article Snippet: The cDNA generated by the PCR reaction described above using SL35-F-(13F) and SL35-R-(13R) was digested with NheI (New England BioLabs Inc.) and cloned into the expression vector pTre2hyg (CLONTECH Laboratories Inc., Palo Alto, California, USA) that had been digested with NheI and EcoRV (New England BioLabs Inc.). .. After amplification in bacteria (TOP10F′; Invitrogen Corp., San Diego, California, USA), the vector constructs were purified, and the sequence of the coding region was verified by direct sequencing using a LI-COR sequencer (LI-COR Biosciences, Bad Homburg, Germany).

    FLAG-tag:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Generated:

    Article Title: A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If)
    Article Snippet: .. The cDNA generated by the PCR reaction described above using SL35-F-(13F) and SL35-R-(13R) was digested with NheI (New England BioLabs Inc.) and cloned into the expression vector pTre2hyg (CLONTECH Laboratories Inc., Palo Alto, California, USA) that had been digested with NheI and EcoRV (New England BioLabs Inc.). .. After amplification in bacteria (TOP10F′; Invitrogen Corp., San Diego, California, USA), the vector constructs were purified, and the sequence of the coding region was verified by direct sequencing using a LI-COR sequencer (LI-COR Biosciences, Bad Homburg, Germany).

    Construct:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Article Title: A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If)
    Article Snippet: The cDNA generated by the PCR reaction described above using SL35-F-(13F) and SL35-R-(13R) was digested with NheI (New England BioLabs Inc.) and cloned into the expression vector pTre2hyg (CLONTECH Laboratories Inc., Palo Alto, California, USA) that had been digested with NheI and EcoRV (New England BioLabs Inc.). .. After amplification in bacteria (TOP10F′; Invitrogen Corp., San Diego, California, USA), the vector constructs were purified, and the sequence of the coding region was verified by direct sequencing using a LI-COR sequencer (LI-COR Biosciences, Bad Homburg, Germany).

    Expressing:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Article Title: A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If)
    Article Snippet: .. The cDNA generated by the PCR reaction described above using SL35-F-(13F) and SL35-R-(13R) was digested with NheI (New England BioLabs Inc.) and cloned into the expression vector pTre2hyg (CLONTECH Laboratories Inc., Palo Alto, California, USA) that had been digested with NheI and EcoRV (New England BioLabs Inc.). .. After amplification in bacteria (TOP10F′; Invitrogen Corp., San Diego, California, USA), the vector constructs were purified, and the sequence of the coding region was verified by direct sequencing using a LI-COR sequencer (LI-COR Biosciences, Bad Homburg, Germany).

    Sequencing:

    Article Title: A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If)
    Article Snippet: The cDNA generated by the PCR reaction described above using SL35-F-(13F) and SL35-R-(13R) was digested with NheI (New England BioLabs Inc.) and cloned into the expression vector pTre2hyg (CLONTECH Laboratories Inc., Palo Alto, California, USA) that had been digested with NheI and EcoRV (New England BioLabs Inc.). .. After amplification in bacteria (TOP10F′; Invitrogen Corp., San Diego, California, USA), the vector constructs were purified, and the sequence of the coding region was verified by direct sequencing using a LI-COR sequencer (LI-COR Biosciences, Bad Homburg, Germany).

    Gel Extraction:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Transformation Assay:

    Article Title: Directed evolution of a picomolar-affinity, high-specificity antibody targeting phosphorylated tau
    Article Snippet: The pThr-231 scFv gene was inserted into the vector pRS316 (a gift from Dr. Eric Shusta), which had been restriction-digested with NheI and BsrGI (New England Biolabs). .. The pRS316 plasmid was transformed into S. cerevisiae YVH10 cells ( ) (a gift from Dr. Eric Shusta) for protein secretion and purification.

    Plasmid Preparation:

    Article Title: The C-Terminal End of HIV-1 Vpu Has a Clade-Specific Determinant That Antagonizes BST-2 and Facilitates Virion Release
    Article Snippet: .. For the Rev-dependent FLAG-tagged MJ4-Vpu expression construct, the vpu coding region from pMJ4 was amplified by PCR with primers designed to introduce a C-terminal FLAG tag, gel purified using a QIAquick gel extraction kit (Qiagen), and then cloned into the pcDNA-RRE expression vector (described in reference ) after digestion of the purified PCR products and backbone with NheI and XhoI (NEB). .. Expression plasmids encoding Vpu-L78A, Vpu-R79A, Vpu-L80A, Vpu-L81A, Vpu L78A,R79A (LR/AA), Vpu-L80A,L81A (LL/AA), Vpu-L78A,R79A,L80A,L81A (LRLL/AAAA), and Vpu-L78H,R79A,L80P,L81W (LRLL/HAPW) with a C-terminal FLAG tag were made by QuikChange site-directed mutagenesis (Agilent) of the MJ4-Vpu-FLAG expression construct.

    Article Title: Directed evolution of a picomolar-affinity, high-specificity antibody targeting phosphorylated tau
    Article Snippet: .. The pThr-231 scFv gene was inserted into the vector pRS316 (a gift from Dr. Eric Shusta), which had been restriction-digested with NheI and BsrGI (New England Biolabs). .. This plasmid included a C-terminal hexahistidine epitope tag.

    Article Title: A mutation in the human MPDU1 gene causes congenital disorder of glycosylation type If (CDG-If)
    Article Snippet: .. The cDNA generated by the PCR reaction described above using SL35-F-(13F) and SL35-R-(13R) was digested with NheI (New England BioLabs Inc.) and cloned into the expression vector pTre2hyg (CLONTECH Laboratories Inc., Palo Alto, California, USA) that had been digested with NheI and EcoRV (New England BioLabs Inc.). .. After amplification in bacteria (TOP10F′; Invitrogen Corp., San Diego, California, USA), the vector constructs were purified, and the sequence of the coding region was verified by direct sequencing using a LI-COR sequencer (LI-COR Biosciences, Bad Homburg, Germany).

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    New England Biolabs nhei
    Electrophoresis of pPrime- fliC Recombinant Plasmids From White Colonies on 1% Agarose Gel Lane 1 to 6: extracted Pprime- fliC plasmids; Lane 8 to 13: Pprime- fliC plasmids digested by <t>XhoI;</t> Lane 15 to 20: Pprime- fliC plasmids digested by XhoI and <t>NheI;</t> Lane 7 and 14: 1 Kbp DNA ladder vivantis.
    Nhei, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nhei/product/New England Biolabs
    Average 99 stars, based on 97 article reviews
    Price from $9.99 to $1999.99
    nhei - by Bioz Stars, 2020-04
    99/100 stars
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    Electrophoresis of pPrime- fliC Recombinant Plasmids From White Colonies on 1% Agarose Gel Lane 1 to 6: extracted Pprime- fliC plasmids; Lane 8 to 13: Pprime- fliC plasmids digested by XhoI; Lane 15 to 20: Pprime- fliC plasmids digested by XhoI and NheI; Lane 7 and 14: 1 Kbp DNA ladder vivantis.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Cloning of fliC Gene From Salmonella typhimurium in the Expression Vector pVAX1 and Evaluation of its Expression in Eukaryotic Cells

    doi: 10.5812/jjm.12351

    Figure Lengend Snippet: Electrophoresis of pPrime- fliC Recombinant Plasmids From White Colonies on 1% Agarose Gel Lane 1 to 6: extracted Pprime- fliC plasmids; Lane 8 to 13: Pprime- fliC plasmids digested by XhoI; Lane 15 to 20: Pprime- fliC plasmids digested by XhoI and NheI; Lane 7 and 14: 1 Kbp DNA ladder vivantis.

    Article Snippet: The recombinant plasmids were detected by restriction digestion with NheI and XhoI enzymes (New England BioLabs, USA) and confirmed by sequence analysis (Bioneer, Korea).

    Techniques: Electrophoresis, Recombinant, Agarose Gel Electrophoresis

    AgaroseGel Electrophoresis of Restriction Enzyme Digestion of Purified pVAX- fliC Recombinant Plasmid Lane 1: pVAX- fliC plasmid digested by XhoI had one band (4419 bp); Lane 2: Double Digestion by NheI and XhoI on pVAX- fliC plasmid had two bands that were 1509 bp (down) and 2910 bp (up); Lane 3: 1 Kbp DNA ladder Vivantis.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Cloning of fliC Gene From Salmonella typhimurium in the Expression Vector pVAX1 and Evaluation of its Expression in Eukaryotic Cells

    doi: 10.5812/jjm.12351

    Figure Lengend Snippet: AgaroseGel Electrophoresis of Restriction Enzyme Digestion of Purified pVAX- fliC Recombinant Plasmid Lane 1: pVAX- fliC plasmid digested by XhoI had one band (4419 bp); Lane 2: Double Digestion by NheI and XhoI on pVAX- fliC plasmid had two bands that were 1509 bp (down) and 2910 bp (up); Lane 3: 1 Kbp DNA ladder Vivantis.

    Article Snippet: The recombinant plasmids were detected by restriction digestion with NheI and XhoI enzymes (New England BioLabs, USA) and confirmed by sequence analysis (Bioneer, Korea).

    Techniques: Electrophoresis, Purification, Recombinant, Plasmid Preparation

    A, Sub-cloning Results of Optimized tPA sp -PADRE-Truncated ORF2 (aa 112 - 660) Gene Cassette In Pvax1 Eukaryotic Plasmid; Lane M, 1kb DNA marker; Lane 1, pBMH-tPA sp -PADRE-truncated ORF2 plasmid; Lane 2, digested pBMH-tPA sp -PADRE-truncated ORF2 plasmid by NheI restriction enzyme; Lane 3, digested pBMH-tPA sp -PADRE-truncated ORF2 plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2900-bp and a 1795-bp fragment; Lane 4, pVAX plasmid; Lane 5, digested pVAX plasmid by NheI and XhoI restriction enzymes and production of 2909-bp band; Lane 6, pVAX-tPA sp -PADRE-truncated ORF2 plasmid; Lane 7, digested pVAX-tPA sp -PADRE-truncated ORF2 plasmid by the NheI restriction enzyme; Lane 8, digested pVAX-tPA sp -PADRE-truncated ORF2 plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2909-bp and a 1795-bp fragment; Lane 9, 1876-bp band in colony PCR test with T7 and BHG universal primers; B, Restriction Enzyme analyses and colony polymerase chain reaction results of three colonies containing pVAX-tPA sp -PADRE-truncated ORF2 plasmid; Lane M, 1 kb DNA marker; Lane 1, negative control; Lanes 2 - 4, 1876-bp band in colony PCR test; Lanes 5 - 7, digested pVAX-tPA sp -PADRE-truncated ORF2 (aa 112 - 660) -linker plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2909-bp and a 1795-bp fragment

    Journal: Jundishapur Journal of Microbiology

    Article Title: Design, Construction and Cloning of Truncated ORF2 and tPAsp-PADRE-Truncated ORF2 Gene Cassette From Hepatitis E Virus in the pVAX1 Expression Vector

    doi: 10.5812/jjm.26035

    Figure Lengend Snippet: A, Sub-cloning Results of Optimized tPA sp -PADRE-Truncated ORF2 (aa 112 - 660) Gene Cassette In Pvax1 Eukaryotic Plasmid; Lane M, 1kb DNA marker; Lane 1, pBMH-tPA sp -PADRE-truncated ORF2 plasmid; Lane 2, digested pBMH-tPA sp -PADRE-truncated ORF2 plasmid by NheI restriction enzyme; Lane 3, digested pBMH-tPA sp -PADRE-truncated ORF2 plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2900-bp and a 1795-bp fragment; Lane 4, pVAX plasmid; Lane 5, digested pVAX plasmid by NheI and XhoI restriction enzymes and production of 2909-bp band; Lane 6, pVAX-tPA sp -PADRE-truncated ORF2 plasmid; Lane 7, digested pVAX-tPA sp -PADRE-truncated ORF2 plasmid by the NheI restriction enzyme; Lane 8, digested pVAX-tPA sp -PADRE-truncated ORF2 plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2909-bp and a 1795-bp fragment; Lane 9, 1876-bp band in colony PCR test with T7 and BHG universal primers; B, Restriction Enzyme analyses and colony polymerase chain reaction results of three colonies containing pVAX-tPA sp -PADRE-truncated ORF2 plasmid; Lane M, 1 kb DNA marker; Lane 1, negative control; Lanes 2 - 4, 1876-bp band in colony PCR test; Lanes 5 - 7, digested pVAX-tPA sp -PADRE-truncated ORF2 (aa 112 - 660) -linker plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2909-bp and a 1795-bp fragment

    Article Snippet: Cloning of Codon-Optimized Truncated ORF2 (aa 112 - 660) Segment in pVAX1 Eukaryotic Expression Vector After cloning of the truncated ORF2 PCR product (without PADRE and tPAsp sequences) in the pVAX1 vector, the recombinant plasmids were detected by colony PCR and restriction enzyme digestion with NheI and XhoI enzymes (New England BioLabs, USA) ( ).

    Techniques: Subcloning, Plasmid Preparation, Marker, Polymerase Chain Reaction, Negative Control

    A, Polymerase Chain Reaction Results of pVAX-Truncated ORF2 (aa 112 - 660) Expression Plasmid (without PADRE and tPA sp sequences); Lane M, 1 kb DNA marker; Lane 1, pVAX-truncated ORF2 (aa 112 - 660) plasmid; Lane 2, digested pVAX-truncated ORF2 (aa 112 - 660) plasmid by NheI restriction enzyme (4596 bp fragment); Lanes 3, digested pVAX-truncated ORF2 (112 - 660) plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2909-bp and a 1693-bp fragment; Lanes 4, PCR product of truncated ORF2 (aa 112-660) gene (without PADRE and tPA sp sequences) with specific primers (1710 bp); Lanes 4, PCR product of tPA sp -PADRE-ORF2 (aa 112 - 660) gene (with PADRE and tPA sp sequences) with T7p and BGH primers (1876 bp); B, Colony Polymerase Chain Reaction Results of Colonies Containing the pVAX-Truncated ORF2 (aa 112 - 660) Expression Recombinant Plasmid; Lane M, 1 kb DNA marker; Lane 1, negative control; Lanes 2 - 9, 1774 bp band of pVAX-truncated ORF2 (aa 112 - 660) plasmid (without PADRE and tPA sp sequences) due to colony PCR assay with T7p and BGH universal primers; Lane 2, 1876 bp band of pVAX-tPA sp -PADRE-ORF2 (aa 112 - 660) plasmid (with PADRE and tPA sp sequences) due to colony PCR assay with T7p and BGH primers as controls

    Journal: Jundishapur Journal of Microbiology

    Article Title: Design, Construction and Cloning of Truncated ORF2 and tPAsp-PADRE-Truncated ORF2 Gene Cassette From Hepatitis E Virus in the pVAX1 Expression Vector

    doi: 10.5812/jjm.26035

    Figure Lengend Snippet: A, Polymerase Chain Reaction Results of pVAX-Truncated ORF2 (aa 112 - 660) Expression Plasmid (without PADRE and tPA sp sequences); Lane M, 1 kb DNA marker; Lane 1, pVAX-truncated ORF2 (aa 112 - 660) plasmid; Lane 2, digested pVAX-truncated ORF2 (aa 112 - 660) plasmid by NheI restriction enzyme (4596 bp fragment); Lanes 3, digested pVAX-truncated ORF2 (112 - 660) plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2909-bp and a 1693-bp fragment; Lanes 4, PCR product of truncated ORF2 (aa 112-660) gene (without PADRE and tPA sp sequences) with specific primers (1710 bp); Lanes 4, PCR product of tPA sp -PADRE-ORF2 (aa 112 - 660) gene (with PADRE and tPA sp sequences) with T7p and BGH primers (1876 bp); B, Colony Polymerase Chain Reaction Results of Colonies Containing the pVAX-Truncated ORF2 (aa 112 - 660) Expression Recombinant Plasmid; Lane M, 1 kb DNA marker; Lane 1, negative control; Lanes 2 - 9, 1774 bp band of pVAX-truncated ORF2 (aa 112 - 660) plasmid (without PADRE and tPA sp sequences) due to colony PCR assay with T7p and BGH universal primers; Lane 2, 1876 bp band of pVAX-tPA sp -PADRE-ORF2 (aa 112 - 660) plasmid (with PADRE and tPA sp sequences) due to colony PCR assay with T7p and BGH primers as controls

    Article Snippet: Cloning of Codon-Optimized Truncated ORF2 (aa 112 - 660) Segment in pVAX1 Eukaryotic Expression Vector After cloning of the truncated ORF2 PCR product (without PADRE and tPAsp sequences) in the pVAX1 vector, the recombinant plasmids were detected by colony PCR and restriction enzyme digestion with NheI and XhoI enzymes (New England BioLabs, USA) ( ).

    Techniques: Polymerase Chain Reaction, Expressing, Plasmid Preparation, Marker, Recombinant, Negative Control

    Vector maps and PEG purification. a Basic vector maps (not to scale) for the original Tet-pLKO-Puro vector and our modified versions. b Agarose gel electrophoresis comparing DNA precipitation methods. 10 μg of EZ-Tet-pLKO vector DNA was co-digested with NheI + EcoRI. The digest was split into three 3 μg aliquots and precipitated with isopropanol (Iso) or polyethylene glycol (PEG) at 6 or 8% concentration. 1 μg of control DNA (uncut and cut) was run alongside 1/3 of the precipitated DNA samples

    Journal: BMC Biotechnology

    Article Title: A streamlined method for the design and cloning of shRNAs into an optimized Dox-inducible lentiviral vector

    doi: 10.1186/s12896-017-0341-x

    Figure Lengend Snippet: Vector maps and PEG purification. a Basic vector maps (not to scale) for the original Tet-pLKO-Puro vector and our modified versions. b Agarose gel electrophoresis comparing DNA precipitation methods. 10 μg of EZ-Tet-pLKO vector DNA was co-digested with NheI + EcoRI. The digest was split into three 3 μg aliquots and precipitated with isopropanol (Iso) or polyethylene glycol (PEG) at 6 or 8% concentration. 1 μg of control DNA (uncut and cut) was run alongside 1/3 of the precipitated DNA samples

    Article Snippet: Vector digest and PEG precipitation Vector can be prepared by co-digesting EZ-Tet-pLKO-Puro DNA with NheI and EcoRI (NEB).

    Techniques: Plasmid Preparation, Purification, Modification, Agarose Gel Electrophoresis, Concentration Assay

    Dendrogram based on Nhe I PFGE patterns of 40 N. gonorrhoeae isolates.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Molecular Basis of High-Level Ciprofloxacin Resistance in Neisseria gonorrhoeae Strains Isolated in Denmark from 1995 to 1998

    doi: 10.1128/AAC.45.1.117-123.2001

    Figure Lengend Snippet: Dendrogram based on Nhe I PFGE patterns of 40 N. gonorrhoeae isolates.

    Article Snippet: Slices of DNA-containing agarose blocks were digested with Spe I and Nhe I (New England Biolabs) overnight at 37°C in 100 μl of restriction endonuclease buffer containing 10 U of enzyme.

    Techniques: