Structured Review

Stratagene puc19
(GAA·TTC) n constructs used to analyze repeat instability. (GAA·TTC) n sequences of the indicated lengths were cloned into the Pst I/Xba I sites of <t>pUC19</t> in both orientations relative to the unidirectional pMB1 origin of replication. Repeat-containing plasmids are depicted in either the ‘GAA’ or ‘TTC’ orientations, based on whether (GAA) n or (TTC) n serves as the lagging strand template, respectively. The plasmid constructs contain repeat lengths of n = 21, 41 and 79. The black boxes flanking the repeat represent minimal flanking sequence from intron 1 of the FXN gene.
Puc19, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/puc19/product/Stratagene
Average 92 stars, based on 28 article reviews
Price from $9.99 to $1999.99
puc19 - by Bioz Stars, 2020-05
92/100 stars

Images

1) Product Images from "Deficiency of RecA-dependent RecFOR and RecBCD pathways causes increased instability of the (GAA?TTC)n sequence when GAA is the lagging strand template"

Article Title: Deficiency of RecA-dependent RecFOR and RecBCD pathways causes increased instability of the (GAA?TTC)n sequence when GAA is the lagging strand template

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkm810

(GAA·TTC) n constructs used to analyze repeat instability. (GAA·TTC) n sequences of the indicated lengths were cloned into the Pst I/Xba I sites of pUC19 in both orientations relative to the unidirectional pMB1 origin of replication. Repeat-containing plasmids are depicted in either the ‘GAA’ or ‘TTC’ orientations, based on whether (GAA) n or (TTC) n serves as the lagging strand template, respectively. The plasmid constructs contain repeat lengths of n = 21, 41 and 79. The black boxes flanking the repeat represent minimal flanking sequence from intron 1 of the FXN gene.
Figure Legend Snippet: (GAA·TTC) n constructs used to analyze repeat instability. (GAA·TTC) n sequences of the indicated lengths were cloned into the Pst I/Xba I sites of pUC19 in both orientations relative to the unidirectional pMB1 origin of replication. Repeat-containing plasmids are depicted in either the ‘GAA’ or ‘TTC’ orientations, based on whether (GAA) n or (TTC) n serves as the lagging strand template, respectively. The plasmid constructs contain repeat lengths of n = 21, 41 and 79. The black boxes flanking the repeat represent minimal flanking sequence from intron 1 of the FXN gene.

Techniques Used: Construct, Clone Assay, Plasmid Preparation, Sequencing

RecA deficiency causes increased instability when GAA is the lagging strand template irrespective of transcription through the repeat tract, or the distance between origin of replication and the (GAA·TTC) 79 sequence. (A) The (GAA·TTC) 79 sequence was additionally subcloned in the GAA orientation into the pDEL (with a deletion of the Plac promoter in pUC19) and pINS (with a 1.5 kb spacer from intron 1 of the human FXN gene inserted in pUC19 between the origin of replication and the repeat tract) vectors (see Materials and Methods section for details). (B) Instability of the (GAA·TTC) 79 sequence was significantly enhanced in the M152 (RecA-deficient) versus MM28 (RecA-proficient) strain. Error bars depict +/− 2SEM; ** P
Figure Legend Snippet: RecA deficiency causes increased instability when GAA is the lagging strand template irrespective of transcription through the repeat tract, or the distance between origin of replication and the (GAA·TTC) 79 sequence. (A) The (GAA·TTC) 79 sequence was additionally subcloned in the GAA orientation into the pDEL (with a deletion of the Plac promoter in pUC19) and pINS (with a 1.5 kb spacer from intron 1 of the human FXN gene inserted in pUC19 between the origin of replication and the repeat tract) vectors (see Materials and Methods section for details). (B) Instability of the (GAA·TTC) 79 sequence was significantly enhanced in the M152 (RecA-deficient) versus MM28 (RecA-proficient) strain. Error bars depict +/− 2SEM; ** P

Techniques Used: Sequencing

Related Articles

Clone Assay:

Article Title: Anisomycin Selectively Desensitizes Signalling Components Involved in Stress Kinase Activation and fos and jun Induction
Article Snippet: .. All other probes were derived from cDNA clones of fos and jun genes, which were generously provided by Rodrigo Bravo (Roche); a 1.7-kb Eco RV/ Hin dIII fragment of fosB in pGEM-1 (Promega), a 0.75-kb Eco RI/ Sac II fragment derived from the 2.5-kb mouse c- jun clone pAH119 ( ) in pUC19, a 1.8-kb Eco RI fragment of junB in pBluescript KS+ (Stratagene), and a 1.5-kb Bam HI/ Hin dIII fragment of junD in pBluescript KS+ (Stratagene). .. All Northern blots were sequentially hybridized to these six probes.

Article Title: Characterization of an unusual tRNA-like sequence found inserted in a Neurospora retroplasmid
Article Snippet: .. The Eco RI-4 fragment of the Hanapepe mtDNA was cloned into pUC19 (Stratagene) and sequenced using the dideoxy chain termination method ( ). .. Sequence comparisons with GenBank were done using the BLAST algorithm with the BLOSUM 62 matrix and an expect value of 10 ( ) at the National Center for Biotechnology Information website.

Article Title: Characterization of an unusual tRNA-like sequence found inserted in a Neurospora retroplasmid
Article Snippet: .. The Eco RI-4 fragment of the Hanapepe mtDNA was cloned into pUC19 (Stratagene) and sequenced using the dideoxy chain termination method ( ). .. Sequence comparisons with GenBank were done using the BLAST algorithm with the BLOSUM 62 matrix and an expect value of 10 ( ) at the National Center for Biotechnology Information website.

Article Title: Role of Two Novel Two-Component Regulatory Systems in Development and Phosphatase Expression in Myxococcus xanthus
Article Snippet: .. E. coli strains were grown at 37°C in LB medium , which was supplemented with ampicillin (50 μg/ml), kanamycin (25 μg/ml), and/or BCIP (40 μg/ml), when needed. pUC19 ( ) and pBluescript SK+ (Stratagene) were used for routine cloning. .. The kanamycin resistance gene was obtained from plasmid pUC7Skm(Pst− ), which was kindly provided by S. Inouye (University of Medicine and Dentistry of New Jersey).

shRNA:

Article Title: The Interaction of CtIP and Nbs1 Connects CDK and ATM to Regulate HR-Mediated Double-Strand Break Repair
Article Snippet: .. Plasmids, mutagenesis, and shRNA/RNAi CtIP cDNA was subcloned into pUC19 at BamHI/SalI and used to generate CtIP variants by site-directed mutagenesis (Stratagene). .. CtIP wild-type and indicated mutants were then subcloned into mammalian expression vectors pcDNA3 or pBabepuro containing HA, Myc or Flag epitopes.

Mutagenesis:

Article Title: Deficiency of RecA-dependent RecFOR and RecBCD pathways causes increased instability of the (GAA?TTC)n sequence when GAA is the lagging strand template
Article Snippet: .. Deletion of the Plac promoter in pUC19, to produce the pDEL-GAA-79 construct ( A), was accomplished by first introducing an Apa I site at the −35 position using the QuikChange II XL site-directed mutagenesis kit (Stratagene), followed by removal of the fragment between Apa I and Hind III, thus deleting both the −10 and −35 sites. ..

Article Title: The Interaction of CtIP and Nbs1 Connects CDK and ATM to Regulate HR-Mediated Double-Strand Break Repair
Article Snippet: .. Plasmids, mutagenesis, and shRNA/RNAi CtIP cDNA was subcloned into pUC19 at BamHI/SalI and used to generate CtIP variants by site-directed mutagenesis (Stratagene). .. CtIP wild-type and indicated mutants were then subcloned into mammalian expression vectors pcDNA3 or pBabepuro containing HA, Myc or Flag epitopes.

Construct:

Article Title: Deficiency of RecA-dependent RecFOR and RecBCD pathways causes increased instability of the (GAA?TTC)n sequence when GAA is the lagging strand template
Article Snippet: .. Deletion of the Plac promoter in pUC19, to produce the pDEL-GAA-79 construct ( A), was accomplished by first introducing an Apa I site at the −35 position using the QuikChange II XL site-directed mutagenesis kit (Stratagene), followed by removal of the fragment between Apa I and Hind III, thus deleting both the −10 and −35 sites. ..

Transformation Assay:

Article Title: The Response Regulator PhoP Is Important for Survival under Conditions of Macrophage-Induced Stress and Virulence in Yersinia pestis
Article Snippet: .. The 262-bp products were digested with Pst I and Hin dIII, ligated into similarly digested pUC19, and transformed in E. coli XL2-Blue MRF′ cells (Stratagene Europe, Amsterdam, The Netherlands). .. The cloned fragments were sequenced using the dideoxynucleotide chain termination method with an Applied Biosystems (Warrington, United Kingdom) PRISM sequencing kit, and the data were compared with other PhoP sequences using BLASTX software ( ).

Derivative Assay:

Article Title: Anisomycin Selectively Desensitizes Signalling Components Involved in Stress Kinase Activation and fos and jun Induction
Article Snippet: .. All other probes were derived from cDNA clones of fos and jun genes, which were generously provided by Rodrigo Bravo (Roche); a 1.7-kb Eco RV/ Hin dIII fragment of fosB in pGEM-1 (Promega), a 0.75-kb Eco RI/ Sac II fragment derived from the 2.5-kb mouse c- jun clone pAH119 ( ) in pUC19, a 1.8-kb Eco RI fragment of junB in pBluescript KS+ (Stratagene), and a 1.5-kb Bam HI/ Hin dIII fragment of junD in pBluescript KS+ (Stratagene). .. All Northern blots were sequentially hybridized to these six probes.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 85
    Stratagene plasmid pfl38 ri
    In vitro binding of wild-type and mutated Arg80 and Arg80-Mcm1-Arg80 proteins to arginine boxes. The end-labeled 160-bp Alu I- Alu I DNA fragment containing the ARG5,6 ). Strain 02463dΔARG80 containing pYEP34 ( ARG81 LEU2 ) was transformed with <t>pFL38</t> (vector; URA3 ) (lane 1), pFL38-RI ( ARG80 ) (lane 2), pAJ234 ( arg80 -Y87F-Y110F) (lane 3), pAJ240 ( arg80- Y110F-T135S) (lane 4), pAJ239 ( arg80 -Y110F-T135S-βI swap) (lane 5), pHL13 ( Arg80-Mcm1-Arg80 ) (lane 6), pAJ46 ( Arg80 - mcm1 -N-terminal/βI/αII swap- Arg80 ) (lane 7), pAJ84 ( Arg80 - mcm1 -N-terminal/αI/βI swap- Arg80 ) (lane 8), and pAJ85 ( Arg80 - mcm1 -N-terminal/αI/αII swap- Arg80 ) (lane 9) . All of these strains were grown on 1% galactose.
    Plasmid Pfl38 Ri, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmid pfl38 ri/product/Stratagene
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    plasmid pfl38 ri - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    90
    Stratagene plasmid puc19
    Amplification of <t>pUC19</t> from colonies and M13mp19 from plaques. Amplification reactions were performed as indicated directly on material picked from a colony or a plaque. Half of the radioactively labeled reaction products were cleaved with Eco RI and both cleaved and uncleaved samples were analyzed by agarose gel electrophoresis. The positions of linear, duplex M13, and pUC19 DNA are indicated.
    Plasmid Puc19, supplied by Stratagene, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmid puc19/product/Stratagene
    Average 90 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    plasmid puc19 - by Bioz Stars, 2020-05
    90/100 stars
      Buy from Supplier

    92
    Stratagene puc19
    (GAA·TTC) n constructs used to analyze repeat instability. (GAA·TTC) n sequences of the indicated lengths were cloned into the Pst I/Xba I sites of <t>pUC19</t> in both orientations relative to the unidirectional pMB1 origin of replication. Repeat-containing plasmids are depicted in either the ‘GAA’ or ‘TTC’ orientations, based on whether (GAA) n or (TTC) n serves as the lagging strand template, respectively. The plasmid constructs contain repeat lengths of n = 21, 41 and 79. The black boxes flanking the repeat represent minimal flanking sequence from intron 1 of the FXN gene.
    Puc19, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/puc19/product/Stratagene
    Average 92 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    puc19 - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    Image Search Results


    In vitro binding of wild-type and mutated Arg80 and Arg80-Mcm1-Arg80 proteins to arginine boxes. The end-labeled 160-bp Alu I- Alu I DNA fragment containing the ARG5,6 ). Strain 02463dΔARG80 containing pYEP34 ( ARG81 LEU2 ) was transformed with pFL38 (vector; URA3 ) (lane 1), pFL38-RI ( ARG80 ) (lane 2), pAJ234 ( arg80 -Y87F-Y110F) (lane 3), pAJ240 ( arg80- Y110F-T135S) (lane 4), pAJ239 ( arg80 -Y110F-T135S-βI swap) (lane 5), pHL13 ( Arg80-Mcm1-Arg80 ) (lane 6), pAJ46 ( Arg80 - mcm1 -N-terminal/βI/αII swap- Arg80 ) (lane 7), pAJ84 ( Arg80 - mcm1 -N-terminal/αI/βI swap- Arg80 ) (lane 8), and pAJ85 ( Arg80 - mcm1 -N-terminal/αI/αII swap- Arg80 ) (lane 9) . All of these strains were grown on 1% galactose.

    Journal: Molecular and Cellular Biology

    Article Title: Swapping Functional Specificity of a MADS Box Protein: Residues Required for Arg80 Regulation of Arginine Metabolism

    doi: 10.1128/MCB.22.16.5741-5752.2002

    Figure Lengend Snippet: In vitro binding of wild-type and mutated Arg80 and Arg80-Mcm1-Arg80 proteins to arginine boxes. The end-labeled 160-bp Alu I- Alu I DNA fragment containing the ARG5,6 ). Strain 02463dΔARG80 containing pYEP34 ( ARG81 LEU2 ) was transformed with pFL38 (vector; URA3 ) (lane 1), pFL38-RI ( ARG80 ) (lane 2), pAJ234 ( arg80 -Y87F-Y110F) (lane 3), pAJ240 ( arg80- Y110F-T135S) (lane 4), pAJ239 ( arg80 -Y110F-T135S-βI swap) (lane 5), pHL13 ( Arg80-Mcm1-Arg80 ) (lane 6), pAJ46 ( Arg80 - mcm1 -N-terminal/βI/αII swap- Arg80 ) (lane 7), pAJ84 ( Arg80 - mcm1 -N-terminal/αI/βI swap- Arg80 ) (lane 8), and pAJ85 ( Arg80 - mcm1 -N-terminal/αI/αII swap- Arg80 ) (lane 9) . All of these strains were grown on 1% galactose.

    Article Snippet: The specific mutations were created by in vitro mutagenesis on double-stranded DNA from plasmid pFL38-RI (pUC19 with ARS4 CEN6 URA3 ARG80 ) ( ) by using the QuikChange site-directed mutagenesis kit from Stratagene (see Fig. for a list of all of the resulting mutated plasmids).

    Techniques: In Vitro, Binding Assay, Labeling, Transformation Assay, Plasmid Preparation

    Amplification of pUC19 from colonies and M13mp19 from plaques. Amplification reactions were performed as indicated directly on material picked from a colony or a plaque. Half of the radioactively labeled reaction products were cleaved with Eco RI and both cleaved and uncleaved samples were analyzed by agarose gel electrophoresis. The positions of linear, duplex M13, and pUC19 DNA are indicated.

    Journal: Genome Research

    Article Title: Rapid Amplification of Plasmid and Phage DNA Using Phi29 DNA Polymerase and Multiply-Primed Rolling Circle Amplification

    doi: 10.1101/gr.180501

    Figure Lengend Snippet: Amplification of pUC19 from colonies and M13mp19 from plaques. Amplification reactions were performed as indicated directly on material picked from a colony or a plaque. Half of the radioactively labeled reaction products were cleaved with Eco RI and both cleaved and uncleaved samples were analyzed by agarose gel electrophoresis. The positions of linear, duplex M13, and pUC19 DNA are indicated.

    Article Snippet: Polyethylene tubing (Intramedic, PE20, 1.09-mm outer diameter) 1 cm in length was stabbed into a colony of Escherichia coli transformed with plasmid pUC19 or a plaque of bacteriophage M13mp19 in a lawn of E. coli (XL1-blue, Stratagene).

    Techniques: Amplification, Labeling, Agarose Gel Electrophoresis

    (GAA·TTC) n constructs used to analyze repeat instability. (GAA·TTC) n sequences of the indicated lengths were cloned into the Pst I/Xba I sites of pUC19 in both orientations relative to the unidirectional pMB1 origin of replication. Repeat-containing plasmids are depicted in either the ‘GAA’ or ‘TTC’ orientations, based on whether (GAA) n or (TTC) n serves as the lagging strand template, respectively. The plasmid constructs contain repeat lengths of n = 21, 41 and 79. The black boxes flanking the repeat represent minimal flanking sequence from intron 1 of the FXN gene.

    Journal: Nucleic Acids Research

    Article Title: Deficiency of RecA-dependent RecFOR and RecBCD pathways causes increased instability of the (GAA?TTC)n sequence when GAA is the lagging strand template

    doi: 10.1093/nar/gkm810

    Figure Lengend Snippet: (GAA·TTC) n constructs used to analyze repeat instability. (GAA·TTC) n sequences of the indicated lengths were cloned into the Pst I/Xba I sites of pUC19 in both orientations relative to the unidirectional pMB1 origin of replication. Repeat-containing plasmids are depicted in either the ‘GAA’ or ‘TTC’ orientations, based on whether (GAA) n or (TTC) n serves as the lagging strand template, respectively. The plasmid constructs contain repeat lengths of n = 21, 41 and 79. The black boxes flanking the repeat represent minimal flanking sequence from intron 1 of the FXN gene.

    Article Snippet: Deletion of the Plac promoter in pUC19, to produce the pDEL-GAA-79 construct ( A), was accomplished by first introducing an Apa I site at the −35 position using the QuikChange II XL site-directed mutagenesis kit (Stratagene), followed by removal of the fragment between Apa I and Hind III, thus deleting both the −10 and −35 sites.

    Techniques: Construct, Clone Assay, Plasmid Preparation, Sequencing

    RecA deficiency causes increased instability when GAA is the lagging strand template irrespective of transcription through the repeat tract, or the distance between origin of replication and the (GAA·TTC) 79 sequence. (A) The (GAA·TTC) 79 sequence was additionally subcloned in the GAA orientation into the pDEL (with a deletion of the Plac promoter in pUC19) and pINS (with a 1.5 kb spacer from intron 1 of the human FXN gene inserted in pUC19 between the origin of replication and the repeat tract) vectors (see Materials and Methods section for details). (B) Instability of the (GAA·TTC) 79 sequence was significantly enhanced in the M152 (RecA-deficient) versus MM28 (RecA-proficient) strain. Error bars depict +/− 2SEM; ** P

    Journal: Nucleic Acids Research

    Article Title: Deficiency of RecA-dependent RecFOR and RecBCD pathways causes increased instability of the (GAA?TTC)n sequence when GAA is the lagging strand template

    doi: 10.1093/nar/gkm810

    Figure Lengend Snippet: RecA deficiency causes increased instability when GAA is the lagging strand template irrespective of transcription through the repeat tract, or the distance between origin of replication and the (GAA·TTC) 79 sequence. (A) The (GAA·TTC) 79 sequence was additionally subcloned in the GAA orientation into the pDEL (with a deletion of the Plac promoter in pUC19) and pINS (with a 1.5 kb spacer from intron 1 of the human FXN gene inserted in pUC19 between the origin of replication and the repeat tract) vectors (see Materials and Methods section for details). (B) Instability of the (GAA·TTC) 79 sequence was significantly enhanced in the M152 (RecA-deficient) versus MM28 (RecA-proficient) strain. Error bars depict +/− 2SEM; ** P

    Article Snippet: Deletion of the Plac promoter in pUC19, to produce the pDEL-GAA-79 construct ( A), was accomplished by first introducing an Apa I site at the −35 position using the QuikChange II XL site-directed mutagenesis kit (Stratagene), followed by removal of the fragment between Apa I and Hind III, thus deleting both the −10 and −35 sites.

    Techniques: Sequencing

    Assortment of single nucleotide polymorphisms in 16 DNA sequences derived from blue colonies transformed with set B recombinants. pUC19-06 derived markers are indicated by upward ticks, while pUC19-07 derived markers are depicted with downward ticks. Crossover events (shaded boxes) are concentrated in the half of the recombinant region closest to the P1 primer. Asterisks indicate those sequences derived from colonies that also appeared to contain the non-recombinant pUC19-07 construct.

    Journal: Nucleic Acids Research

    Article Title: Offset recombinant PCR: a simple but effective method for shuffling compact heterologous domains

    doi: 10.1093/nar/gni081

    Figure Lengend Snippet: Assortment of single nucleotide polymorphisms in 16 DNA sequences derived from blue colonies transformed with set B recombinants. pUC19-06 derived markers are indicated by upward ticks, while pUC19-07 derived markers are depicted with downward ticks. Crossover events (shaded boxes) are concentrated in the half of the recombinant region closest to the P1 primer. Asterisks indicate those sequences derived from colonies that also appeared to contain the non-recombinant pUC19-07 construct.

    Article Snippet: pUC19 mutants The QuickChange™ Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA) was used to insert ochre codons and silent point mutation into the N-terminal region of the lacZ (open reading frame) ORF on the pUC19 plasmid (GenBank accession no. L09137 X02514 ).

    Techniques: Derivative Assay, Transformation Assay, Recombinant, Construct

    Design of the lacZ reporter system. ( A ) Point mutations for each of the pUC19 constructs used in our PCR recombination experiments. Although all nucleotide sequences in this paper are presented in terms of the lacZ coding strand, they are numbered in accordance with their compliments for consistency with the published pUC19 sequence. Nucleic acid mutations that lead to the creation of ochre stop codons in the lacZ ORF are shown in bold. All other mutations have no impact on the wild-type β-galactosidase amino acid sequence, which is aligned with the nucleotide sequences at the bottom of the frame. ( B ) Locations of primers, restriction sites and the 82 bp recombinant region (shaded box) with respect to the lacZ ORF on the pUC19 vector. Dashed lines mark center points for the 329 and 511 bp PCR products. Half arrows represent the arrangements of forward and reverse primers (P1, P2 and P3) on the two amplicons.

    Journal: Nucleic Acids Research

    Article Title: Offset recombinant PCR: a simple but effective method for shuffling compact heterologous domains

    doi: 10.1093/nar/gni081

    Figure Lengend Snippet: Design of the lacZ reporter system. ( A ) Point mutations for each of the pUC19 constructs used in our PCR recombination experiments. Although all nucleotide sequences in this paper are presented in terms of the lacZ coding strand, they are numbered in accordance with their compliments for consistency with the published pUC19 sequence. Nucleic acid mutations that lead to the creation of ochre stop codons in the lacZ ORF are shown in bold. All other mutations have no impact on the wild-type β-galactosidase amino acid sequence, which is aligned with the nucleotide sequences at the bottom of the frame. ( B ) Locations of primers, restriction sites and the 82 bp recombinant region (shaded box) with respect to the lacZ ORF on the pUC19 vector. Dashed lines mark center points for the 329 and 511 bp PCR products. Half arrows represent the arrangements of forward and reverse primers (P1, P2 and P3) on the two amplicons.

    Article Snippet: pUC19 mutants The QuickChange™ Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA) was used to insert ochre codons and silent point mutation into the N-terminal region of the lacZ (open reading frame) ORF on the pUC19 plasmid (GenBank accession no. L09137 X02514 ).

    Techniques: Construct, Polymerase Chain Reaction, Sequencing, Recombinant, Plasmid Preparation

    Assortments of single nucleotide polymorphisms in set B recombinants derived from blue ( A ) and white ( B ) colonies after six consecutive rounds of OR-PCR. Purified plasmid samples were retransformed into E.coli before being prepared for sequencing in order to avoid multiple alleles in a single sample. pUC19-06 derived markers are indicated by upward ticks, while pUC19-07 derived markers are depicted with downward ticks. Nucleotide mutations that lead to the creation of ochre stop codons and loss of the lacZ phenotype are represented with gray ticks. The locations of crossover events are marked by shaded boxes.

    Journal: Nucleic Acids Research

    Article Title: Offset recombinant PCR: a simple but effective method for shuffling compact heterologous domains

    doi: 10.1093/nar/gni081

    Figure Lengend Snippet: Assortments of single nucleotide polymorphisms in set B recombinants derived from blue ( A ) and white ( B ) colonies after six consecutive rounds of OR-PCR. Purified plasmid samples were retransformed into E.coli before being prepared for sequencing in order to avoid multiple alleles in a single sample. pUC19-06 derived markers are indicated by upward ticks, while pUC19-07 derived markers are depicted with downward ticks. Nucleotide mutations that lead to the creation of ochre stop codons and loss of the lacZ phenotype are represented with gray ticks. The locations of crossover events are marked by shaded boxes.

    Article Snippet: pUC19 mutants The QuickChange™ Site-Directed Mutagenesis Kit (Stratagene, La Jolla, CA) was used to insert ochre codons and silent point mutation into the N-terminal region of the lacZ (open reading frame) ORF on the pUC19 plasmid (GenBank accession no. L09137 X02514 ).

    Techniques: Derivative Assay, Polymerase Chain Reaction, Purification, Plasmid Preparation, Sequencing