Nru I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 1 article reviews
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1) Product Images from "Mice Devoid of Fer Protein-Tyrosine Kinase Activity Are Viable and Fertile but Display Reduced Cortactin Phosphorylation"
Article Title: Mice Devoid of Fer Protein-Tyrosine Kinase Activity Are Viable and Fertile but Display Reduced Cortactin Phosphorylation
Journal: Molecular and Cellular Biology
Figure Legend Snippet: Domain structure of Fer and targeting strategy of fer locus. (A) Fer consists of three CC motifs, a central SH2 domain, and a kinase domain. The catalytic subdomains are shown, with subdomain IX encoded by exon 19 contained within genomic clone 3B. The targeting vector contains a 6-kbp long arm of homology, followed by the PGK-neo cassette, a 0.9-kbp short arm of homology, and a PGK-tk cassette. The D743R mutation was generated within exon 19, which also introduces an Nru I (N) site. A schematic of the targeted allele following homologous recombination is shown, with positions of PCR primers and Southern blot probe indicated. Positions of Xba I (X), Xho I (Xh), Nhe I (Nh), Eco RI (E), and Sac I (S) sites are shown. Sites in parentheses were destroyed. (B) Southern blot analysis of Sac I-digested genomic DNA from animals that were wild type (+/+), heterozygous (+/−), or homozygous (−/−) for fer D743R . Hybridization with a probe located 3′ to the sequence used for the short arm of homology reveals bands of 4.0 kbp for the wild-type allele and 5.8 kbp for the fer D743R allele. (C) Genotyping PCR analysis of genomic DNA from +/+, +/−, or −/− animals. The resulting 871-bp fragment is resistant to Nru I digestion in wild-type samples, yields additional 605- and 266-bp bands in heterozygous samples, and yields only 605- and 266-bp fragments in homozygous samples.
Techniques Used: Plasmid Preparation, Mutagenesis, Generated, Homologous Recombination, Polymerase Chain Reaction, Southern Blot, Hybridization, Sequencing
2) Product Images from "A Novel Plasmid DNA-Based Foot and Mouth Disease Virus Minigenome for Intracytoplasmic mRNA Production"
Article Title: A Novel Plasmid DNA-Based Foot and Mouth Disease Virus Minigenome for Intracytoplasmic mRNA Production
Figure Legend Snippet: Construction of recombinant plasmids. ( A ) The DNA cassette containing T7 promoter and FMDV O189 5′UTR in pKLS1 vector was removed by digesting with the restriction enzymes, Nru I and Stu I, and placed upstream of FMDV O189 3′UTR, in plasmid pUC57_3′UTR, resulting in the pKLS3 vector. The complete pKLS3 construct is an FMDV minigenome containing the essential FMDV cis-acting elements for viral replication/transcription and translation and a Stu I site for foreign gene insertion. ( B ) The enhanced green fluorescence protein (GFP) gene was inserted into pKLS3 at the Stu I site between FMDV 5′and 3′UTRs to generate a minigenome with a transfection reporter pKLS3_GFP.
Techniques Used: Recombinant, Plasmid Preparation, Construct, Fluorescence, Transfection
3) Product Images from "Isolation of Chlamydia pneumoniae Clonal Variants by a Focus-Forming Assay "
Article Title: Isolation of Chlamydia pneumoniae Clonal Variants by a Focus-Forming Assay
Journal: Infection and Immunity
Figure Legend Snippet: (A) Genomic map of the tyrP region. Sequencing data revealed a 1,649-bp polymorphism containing a duplication of the tyrP and ycc ). Lines indicate the cutting sites of Nru I resulting in 3,974- and 5,623-bp fragments, respectively. The bar indicates the region amplified by the duplication-PCR. (B) Southern blot analysis after Nru I digestion. The MUL-250 wild-type strain shows two bands of 3,974 and 5,623 bp, respectively, indicating the presence of both a single tyrP copy population and a double tyrP copy population. Clones obtained by the focus assay showed either a double copy (clone 38) or a single copy (clone 41). HeLa DNA was used as a negative control (line 4). (C) Agarose gel analysis of duplication-PCR products (93 bp). Screening of 60 C. pneumoniae strain MUL-250 clones revealed a single clone (clone 38) containing a duplicated tyrP gene as indicated by a positive PCR product. A negative duplication-PCR for clone 41 indicates the presence of a single tyrP copy only. The MUL-250 wild-type strain was used as a positive control, and H 2 O was used as a negative control (lane 4).
Techniques Used: Sequencing, Amplification, Polymerase Chain Reaction, Southern Blot, Clone Assay, Negative Control, Agarose Gel Electrophoresis, Positive Control
4) Product Images from "Comparative genomics of drug resistance in Trypanosoma brucei rhodesiense"
Article Title: Comparative genomics of drug resistance in Trypanosoma brucei rhodesiense
Journal: Cellular and Molecular Life Sciences
Figure Legend Snippet: Loss or mutation of AT1 in the resistant lines . a Genomic 454 reads of STIB900-P and STIB900-M mapped to STIB900 visualized for the AT1 locus on chromosome 5 using BamView [ 59 ] (smaller scale for STIB900-M). There is a deletion of AT1 in STIB900-M and a coding point mutation in STIB900-P ( red ). b AT1 PCR products (1636 bp) were amplified from genomic DNA of STIB900 and STIB900-P, and digested with the endonuclease Nru I. G1288C mutant alleles are cut to fragments of 1339 and 257 bp. STIB900-P appears to be heterozygous for the mutation
Techniques Used: Mutagenesis, Polymerase Chain Reaction, Amplification