r0131  (New England Biolabs)

Journal: Journal of Clinical Microbiology

Article Title: Molecular Characterization of Quinolone-Resistant Neisseria gonorrhoeae Isolates from Brazil ▿

doi: 10.1128/JCM.01175-11

Figure Lengend Snippet: Dendrogram based on NheI PFGE patterns of the genomic DNA of 25 ciprofloxacin-resistant N. gonorrhoeae isolates from Rio de Janeiro, Brazil. Abbreviations: PPNG, penicillinase-producing N. gonorrhoeae ; TRNG, plasmid-mediated tetracycline resistance; CMRP,

Article Snippet: Subsequently, each DNA preparation was digested with NheI (New England Biolabs, Ipswich, MA) overnight at 37°C in 100 μl of restriction endonuclease buffer containing 15 U of the enzyme.

Techniques: Plasmid Preparation

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

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

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

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

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