spei  (New England Biolabs)


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    Structured Review

    New England Biolabs spei
    OSN-specific GABAR B1 and D2R knockout mice, Related to Figure 2 . (A) Gene targeting at Drd2 locus. We obtained germline transmission from one ES clone, HEPD064_5_D11. The Drd2 locus in Drd2 tm1a , Drd2 fl (= Drd tm1c ) and Drd cKO . Drd2 +/ tm1a mice were crossed with Flp mice to obtain Drd2 +/ fl . ( C ) Gene targeting at Gabbr1 locus. We obtained germline transmission from one ES clone, EPD0730_1_G04. ( D ) The Gabbr1 locus in Gabbr1 tm1a , Gabbr1 fl (= Gabbr1 tm1c ), and Gabbr1 cKO . Gabbr1 +/ tm1a mice were crossed with Flp mice to obtain Gabbr1 +/ fl . ( E-G ) Southern blot analysis on Drd2 +/+ and Drd2 tm1a /+ ( E , Drd2 probe), Gabbr1 +/+ and Gabbr1 tm1a /+ ( F , Gabbr1 probe); Drd2 tm1a /+ and Gabbr1 tm1a /+ ( G , neo r probe). ( H ) Immunostaining of D2R in the OB of OSN-specific Drd2 mutant mice. ( I ) Immunostaining of GABAR B1 in the OB of OSN-specific Gabbr1 mutant mice. Location of 5’ and 3’ arms for gene targeting and 5’ and 3’ DNA probes for southern blotting are shown in ( A ) and ( C ). <t>KpnI</t> (K), <t>SpeI</t> (S), XhoI (X) and ApaI (A).
    Spei, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Widespread inhibition, antagonism, and synergy in mouse olfactory sensory neurons in vivo"

    Article Title: Widespread inhibition, antagonism, and synergy in mouse olfactory sensory neurons in vivo

    Journal: bioRxiv

    doi: 10.1101/803908

    OSN-specific GABAR B1 and D2R knockout mice, Related to Figure 2 . (A) Gene targeting at Drd2 locus. We obtained germline transmission from one ES clone, HEPD064_5_D11. The Drd2 locus in Drd2 tm1a , Drd2 fl (= Drd tm1c ) and Drd cKO . Drd2 +/ tm1a mice were crossed with Flp mice to obtain Drd2 +/ fl . ( C ) Gene targeting at Gabbr1 locus. We obtained germline transmission from one ES clone, EPD0730_1_G04. ( D ) The Gabbr1 locus in Gabbr1 tm1a , Gabbr1 fl (= Gabbr1 tm1c ), and Gabbr1 cKO . Gabbr1 +/ tm1a mice were crossed with Flp mice to obtain Gabbr1 +/ fl . ( E-G ) Southern blot analysis on Drd2 +/+ and Drd2 tm1a /+ ( E , Drd2 probe), Gabbr1 +/+ and Gabbr1 tm1a /+ ( F , Gabbr1 probe); Drd2 tm1a /+ and Gabbr1 tm1a /+ ( G , neo r probe). ( H ) Immunostaining of D2R in the OB of OSN-specific Drd2 mutant mice. ( I ) Immunostaining of GABAR B1 in the OB of OSN-specific Gabbr1 mutant mice. Location of 5’ and 3’ arms for gene targeting and 5’ and 3’ DNA probes for southern blotting are shown in ( A ) and ( C ). KpnI (K), SpeI (S), XhoI (X) and ApaI (A).
    Figure Legend Snippet: OSN-specific GABAR B1 and D2R knockout mice, Related to Figure 2 . (A) Gene targeting at Drd2 locus. We obtained germline transmission from one ES clone, HEPD064_5_D11. The Drd2 locus in Drd2 tm1a , Drd2 fl (= Drd tm1c ) and Drd cKO . Drd2 +/ tm1a mice were crossed with Flp mice to obtain Drd2 +/ fl . ( C ) Gene targeting at Gabbr1 locus. We obtained germline transmission from one ES clone, EPD0730_1_G04. ( D ) The Gabbr1 locus in Gabbr1 tm1a , Gabbr1 fl (= Gabbr1 tm1c ), and Gabbr1 cKO . Gabbr1 +/ tm1a mice were crossed with Flp mice to obtain Gabbr1 +/ fl . ( E-G ) Southern blot analysis on Drd2 +/+ and Drd2 tm1a /+ ( E , Drd2 probe), Gabbr1 +/+ and Gabbr1 tm1a /+ ( F , Gabbr1 probe); Drd2 tm1a /+ and Gabbr1 tm1a /+ ( G , neo r probe). ( H ) Immunostaining of D2R in the OB of OSN-specific Drd2 mutant mice. ( I ) Immunostaining of GABAR B1 in the OB of OSN-specific Gabbr1 mutant mice. Location of 5’ and 3’ arms for gene targeting and 5’ and 3’ DNA probes for southern blotting are shown in ( A ) and ( C ). KpnI (K), SpeI (S), XhoI (X) and ApaI (A).

    Techniques Used: Knock-Out, Mouse Assay, Transmission Assay, Southern Blot, Immunostaining, Mutagenesis

    2) Product Images from "Production of enterodiol from defatted flaxseeds through biotransformation by human intestinal bacteria"

    Article Title: Production of enterodiol from defatted flaxseeds through biotransformation by human intestinal bacteria

    Journal: BMC Microbiology

    doi: 10.1186/1471-2180-10-115

    PFGE patterns of SpeI-cleaved genomic DNA of 32 pure cultures obtained from END-49 . Assignment of the bacterial strains to Genome Group I, II, III, IV or V was indicated at the bottom of the PFGE photo.
    Figure Legend Snippet: PFGE patterns of SpeI-cleaved genomic DNA of 32 pure cultures obtained from END-49 . Assignment of the bacterial strains to Genome Group I, II, III, IV or V was indicated at the bottom of the PFGE photo.

    Techniques Used:

    3) Product Images from "Simultaneous Detection and Mutation Surveillance of SARS-CoV-2 and co-infections of multiple respiratory viruses by Rapid field-deployable sequencing"

    Article Title: Simultaneous Detection and Mutation Surveillance of SARS-CoV-2 and co-infections of multiple respiratory viruses by Rapid field-deployable sequencing

    Journal: medRxiv

    doi: 10.1101/2020.06.12.20129247

    Agarose gel electrophoresis results of singleplex RPA a , Agarose gel electrophoresis results of singleplex RPA with selected primers shown next a molecular size marker. The amplicons range from 194 bp to 466 bp. b , Agarose gel electrophoresis results of restriction enzyme digestion. The amplicon of pair 5 was digested by SpeI while the others were digested by NlaIII. The digested DNA bands (asterisks) were of expected sizes. c , Agarose gel electrophoresis results showing the sensitivity of RPA in amplifying the SARS-CoV-2 genome. Primer pair 4 was used in the experiment. Reliable amplification can be achieved with 1.4 copies (calculated from dilution) of the SARS-CoV-2 genome. d , Agarose gel electrophoresis result of one-pot reverse transcription and RPA reaction using primer pair 4.
    Figure Legend Snippet: Agarose gel electrophoresis results of singleplex RPA a , Agarose gel electrophoresis results of singleplex RPA with selected primers shown next a molecular size marker. The amplicons range from 194 bp to 466 bp. b , Agarose gel electrophoresis results of restriction enzyme digestion. The amplicon of pair 5 was digested by SpeI while the others were digested by NlaIII. The digested DNA bands (asterisks) were of expected sizes. c , Agarose gel electrophoresis results showing the sensitivity of RPA in amplifying the SARS-CoV-2 genome. Primer pair 4 was used in the experiment. Reliable amplification can be achieved with 1.4 copies (calculated from dilution) of the SARS-CoV-2 genome. d , Agarose gel electrophoresis result of one-pot reverse transcription and RPA reaction using primer pair 4.

    Techniques Used: Agarose Gel Electrophoresis, Recombinase Polymerase Amplification, Marker, Amplification

    4) Product Images from "Seven novel mutations in the long isoform of the USH2A gene in Chinese families with nonsyndromic retinitis pigmentosa and Usher syndrome Type II"

    Article Title: Seven novel mutations in the long isoform of the USH2A gene in Chinese families with nonsyndromic retinitis pigmentosa and Usher syndrome Type II

    Journal: Molecular Vision

    doi:

    A restriction fragment length analysis of the four mutations detected in this study. A : c.2802T > G abolished a HincII restriction site that co-segregated with the affected individuals and the carriers (42 bp, 57 bp, 99 bp, 717 bp, and 774 bp), but not with unaffected individuals and normal controls (42 bp, 57 bp, and 717 bp). B : c.8232G > C created a new HpyCH4V restriction site that co-segregated with the affected individuals and the carriers (88 bp, 186 bp, 218 bp, and 274 bp), but not with unaffected individuals and normal controls (218 bp, 274 bp). C : c.3788G > A abolished a BsaI restriction site that co-segregated with the affected individuals and the carriers (70 bp, 132 bp, 422 bp, and 492 bp), but not with unaffected individuals and normal controls (70 bp, 132 bp, and 422 bp). D : c.14403C > G created a SpeI restriction site that co-segregated with the affected individuals and the carriers (145 bp, 300 bp, and 445 bp), but not with unaffected individuals and normal controls (445 bp). A participant identification number is given above each lane. N represents normal controls.
    Figure Legend Snippet: A restriction fragment length analysis of the four mutations detected in this study. A : c.2802T > G abolished a HincII restriction site that co-segregated with the affected individuals and the carriers (42 bp, 57 bp, 99 bp, 717 bp, and 774 bp), but not with unaffected individuals and normal controls (42 bp, 57 bp, and 717 bp). B : c.8232G > C created a new HpyCH4V restriction site that co-segregated with the affected individuals and the carriers (88 bp, 186 bp, 218 bp, and 274 bp), but not with unaffected individuals and normal controls (218 bp, 274 bp). C : c.3788G > A abolished a BsaI restriction site that co-segregated with the affected individuals and the carriers (70 bp, 132 bp, 422 bp, and 492 bp), but not with unaffected individuals and normal controls (70 bp, 132 bp, and 422 bp). D : c.14403C > G created a SpeI restriction site that co-segregated with the affected individuals and the carriers (145 bp, 300 bp, and 445 bp), but not with unaffected individuals and normal controls (445 bp). A participant identification number is given above each lane. N represents normal controls.

    Techniques Used:

    5) Product Images from "Molecular Epidemiologic Analysis and Antimicrobial Resistance of Helicobacter cinaedi Isolated from Seven Hospitals in Japan"

    Article Title: Molecular Epidemiologic Analysis and Antimicrobial Resistance of Helicobacter cinaedi Isolated from Seven Hospitals in Japan

    Journal: Journal of Clinical Microbiology

    doi: 10.1128/JCM.06810-11

    Pulsed-field gel electrophoresis patterns of XhoI-digested (A) and SpeI-digested (B) H. cinaedi isolates from Japan as well as the six reference strains. The letter in each isolate's identifier indicates the hospital at which the strain was isolated (e.g.,
    Figure Legend Snippet: Pulsed-field gel electrophoresis patterns of XhoI-digested (A) and SpeI-digested (B) H. cinaedi isolates from Japan as well as the six reference strains. The letter in each isolate's identifier indicates the hospital at which the strain was isolated (e.g.,

    Techniques Used: Pulsed-Field Gel, Electrophoresis, Isolation

    6) Product Images from "Rapid Microarray-Based Genotyping of Enterohemorrhagic Escherichia coli Serotype O156:H25/H-/Hnt Isolates from Cattle and Clonal Relationship Analysis ▿ Serotype O156:H25/H-/Hnt Isolates from Cattle and Clonal Relationship Analysis ▿ †"

    Article Title: Rapid Microarray-Based Genotyping of Enterohemorrhagic Escherichia coli Serotype O156:H25/H-/Hnt Isolates from Cattle and Clonal Relationship Analysis ▿ Serotype O156:H25/H-/Hnt Isolates from Cattle and Clonal Relationship Analysis ▿ †

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.00743-10

    Neighbor-joining tree of bovine E. coli O156 isolates based on the restriction pattern obtained after digestion with XbaI, NotI, BlnI, and SpeI (see Fig. S1 in the supplemental material).
    Figure Legend Snippet: Neighbor-joining tree of bovine E. coli O156 isolates based on the restriction pattern obtained after digestion with XbaI, NotI, BlnI, and SpeI (see Fig. S1 in the supplemental material).

    Techniques Used:

    7) Product Images from "Custom-Designed Molecular Scissors for Site-Specific Manipulation of the Plant and Mammalian Genomes"

    Article Title: Custom-Designed Molecular Scissors for Site-Specific Manipulation of the Plant and Mammalian Genomes

    Journal: Methods in molecular biology (Clifton, N.J.)

    doi: 10.1007/978-1-59745-483-4_40

    Converting ZFPs into ZFNs. The NdeI/SpeI-cut ZFPs are ligated into the pET15b:N, the plasmid containing the FokI cleavage domain to form pET15b:ZFN.
    Figure Legend Snippet: Converting ZFPs into ZFNs. The NdeI/SpeI-cut ZFPs are ligated into the pET15b:N, the plasmid containing the FokI cleavage domain to form pET15b:ZFN.

    Techniques Used: Plasmid Preparation

    8) Product Images from "Gluconobacter as Well as Asaia Species, Newly Emerging Opportunistic Human Pathogens among Acetic Acid Bacteria ▿ Species, Newly Emerging Opportunistic Human Pathogens among Acetic Acid Bacteria ▿ †"

    Article Title: Gluconobacter as Well as Asaia Species, Newly Emerging Opportunistic Human Pathogens among Acetic Acid Bacteria ▿ Species, Newly Emerging Opportunistic Human Pathogens among Acetic Acid Bacteria ▿ †

    Journal: Journal of Clinical Microbiology

    doi: 10.1128/JCM.00767-10

    PFGE patterns of SpeI-restricted DNAs for the five Gluconobacter sp. strains reported in this study. Lanes 1, 2, and 3, strains aP78, aP81, and aP112 (case 2), respectively; lane 4, strain aP90 (case 3); lane 5, strain LBN 175 (case 1); lanes Sc and
    Figure Legend Snippet: PFGE patterns of SpeI-restricted DNAs for the five Gluconobacter sp. strains reported in this study. Lanes 1, 2, and 3, strains aP78, aP81, and aP112 (case 2), respectively; lane 4, strain aP90 (case 3); lane 5, strain LBN 175 (case 1); lanes Sc and

    Techniques Used:

    9) Product Images from "Increased PHGDH expression uncouples hair follicle cycle progression and promotes inappropriate melanin accumulation"

    Article Title: Increased PHGDH expression uncouples hair follicle cycle progression and promotes inappropriate melanin accumulation

    Journal: bioRxiv

    doi: 10.1101/249250

    Generation of the PHGDH tetO allele (A) Schematic of the Col1A locus in wildtype mouse cells (top), the modified locus in KH2 ES cells (middle) and the locus after targeting to introduce the PHGDH tetO allele (bottom). The PHGDH cDNA introduced into the Col1A locus is the human sequence. The expected band sizes when the indicated probe is used for Southern blot analysis as in (B) and (C) ] are indicated. Also shown are the location of the SpeI sites (marked “S”) in each locus used to digest genomic DNA for Southern blot analysis. FRT, flippase recognition target site; tetO, tetracycline operator minimal promoter. (B) Southern blot analysis of SpeI-digested genomic DNA from six PHGDH tetO -targeted ES cells. Clones D4 and D5 exhibit proper targeting of the Col1A locus and an unaffected wild-type allele. (C) Southern blot analysis of SpeI-digested genomic DNA from mice of the indicated genotypes. (D) PCR-based genotyping of the PHGDH tetO and Rosa26-M2rtTA alleles. In the PHGDH tetO reaction, the presence of the transgene is indicated by the upper band; in the Rosa26-M2rtTA reaction, the presence of the transgene is indicated by the lower band. (E) The number of offspring of each genotype observed when mice hemizygous for the PHGDH tetO allele exposed to a doxycycline diet were mated. The observed distribution of genotypes in the offspring did not differ significantly from expected Mendelian ratios, with p=0.58 by the χ 2 goodness-of-fit test. (F) Western blot analysis of PHGDH protein using the indicated amount of recombinant human or mouse PHGDH to test antibody specificity.
    Figure Legend Snippet: Generation of the PHGDH tetO allele (A) Schematic of the Col1A locus in wildtype mouse cells (top), the modified locus in KH2 ES cells (middle) and the locus after targeting to introduce the PHGDH tetO allele (bottom). The PHGDH cDNA introduced into the Col1A locus is the human sequence. The expected band sizes when the indicated probe is used for Southern blot analysis as in (B) and (C) ] are indicated. Also shown are the location of the SpeI sites (marked “S”) in each locus used to digest genomic DNA for Southern blot analysis. FRT, flippase recognition target site; tetO, tetracycline operator minimal promoter. (B) Southern blot analysis of SpeI-digested genomic DNA from six PHGDH tetO -targeted ES cells. Clones D4 and D5 exhibit proper targeting of the Col1A locus and an unaffected wild-type allele. (C) Southern blot analysis of SpeI-digested genomic DNA from mice of the indicated genotypes. (D) PCR-based genotyping of the PHGDH tetO and Rosa26-M2rtTA alleles. In the PHGDH tetO reaction, the presence of the transgene is indicated by the upper band; in the Rosa26-M2rtTA reaction, the presence of the transgene is indicated by the lower band. (E) The number of offspring of each genotype observed when mice hemizygous for the PHGDH tetO allele exposed to a doxycycline diet were mated. The observed distribution of genotypes in the offspring did not differ significantly from expected Mendelian ratios, with p=0.58 by the χ 2 goodness-of-fit test. (F) Western blot analysis of PHGDH protein using the indicated amount of recombinant human or mouse PHGDH to test antibody specificity.

    Techniques Used: Modification, Introduce, Sequencing, Southern Blot, Clone Assay, Mouse Assay, Polymerase Chain Reaction, Genotyping Assay, Western Blot, Recombinant

    10) Product Images from "Defining natural species of bacteria: clear-cut genomic boundaries revealed by a turning point in nucleotide sequence divergence"

    Article Title: Defining natural species of bacteria: clear-cut genomic boundaries revealed by a turning point in nucleotide sequence divergence

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-14-489

    Genomic DNA PFGE patterns of representative Salmonella strains, cleaved with I-CeuI (a), XbaI (b) and SpeI (c), respectively. Lanes: 1, DNA size Marker; 2-5, S. enteritidis (SE310, SE154, SE301, LK5); 6-14, S. pullorum (RKS5078, CDC1983-67, SARB51, 04–6767, NS387, 00–19557, 02–15951, 03–16062, 98–13777); 15–20, S. gallinarum (287/91, RKS5021, SGSC2293, 91–29327, 90–5289, 92–7995).
    Figure Legend Snippet: Genomic DNA PFGE patterns of representative Salmonella strains, cleaved with I-CeuI (a), XbaI (b) and SpeI (c), respectively. Lanes: 1, DNA size Marker; 2-5, S. enteritidis (SE310, SE154, SE301, LK5); 6-14, S. pullorum (RKS5078, CDC1983-67, SARB51, 04–6767, NS387, 00–19557, 02–15951, 03–16062, 98–13777); 15–20, S. gallinarum (287/91, RKS5021, SGSC2293, 91–29327, 90–5289, 92–7995).

    Techniques Used: Marker

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    New England Biolabs spei
    OSN-specific GABAR B1 and D2R knockout mice, Related to Figure 2 . (A) Gene targeting at Drd2 locus. We obtained germline transmission from one ES clone, HEPD064_5_D11. The Drd2 locus in Drd2 tm1a , Drd2 fl (= Drd tm1c ) and Drd cKO . Drd2 +/ tm1a mice were crossed with Flp mice to obtain Drd2 +/ fl . ( C ) Gene targeting at Gabbr1 locus. We obtained germline transmission from one ES clone, EPD0730_1_G04. ( D ) The Gabbr1 locus in Gabbr1 tm1a , Gabbr1 fl (= Gabbr1 tm1c ), and Gabbr1 cKO . Gabbr1 +/ tm1a mice were crossed with Flp mice to obtain Gabbr1 +/ fl . ( E-G ) Southern blot analysis on Drd2 +/+ and Drd2 tm1a /+ ( E , Drd2 probe), Gabbr1 +/+ and Gabbr1 tm1a /+ ( F , Gabbr1 probe); Drd2 tm1a /+ and Gabbr1 tm1a /+ ( G , neo r probe). ( H ) Immunostaining of D2R in the OB of OSN-specific Drd2 mutant mice. ( I ) Immunostaining of GABAR B1 in the OB of OSN-specific Gabbr1 mutant mice. Location of 5’ and 3’ arms for gene targeting and 5’ and 3’ DNA probes for southern blotting are shown in ( A ) and ( C ). <t>KpnI</t> (K), <t>SpeI</t> (S), XhoI (X) and ApaI (A).
    Spei, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/spei/product/New England Biolabs
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    spei - by Bioz Stars, 2022-07
    97/100 stars
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    98
    New England Biolabs spei restriction enzymes
    Plasmid stability. GFP expression was induced with 500 ng/ml of ATc. ON culture of day 0 was supplemented with 15 μg/ml kanamycin, while ON cultures of days 1–4 were supplemented with 50 μg/ml ampicillin. (A) GFP expression in F. novicida U112 pFNMB1 msfgfp ON cultures of day 0 and day 4. (B) GFP expression in F. novicida U112 pFNMB2 msfgfp ON cultures of day 0 and day 4. (A,B) Images are a merge of phase contrast and GFP channel. 26 × 39 μm fields of view are shown. Scale bar represent 5 μm. Representative replicates are shown. Three independent experiments were performed. (C) Survival assay performed with ON cultures of F. novicida U112 pFNMB1 msfgfp and F. novicida U112 pFNMB2 msfgfp plated on ampicillin and ampicillin/kanamycin plates. Three independent experiments were performed. (D) Digestion of pFNMB1 msfgfp with <t>SacI</t> and <t>SpeI</t> restriction enzymes. (E) Digestion of pFNMB2 msfgfp with SacI and SpeI restriction enzymes. (D,E) Plasmids were passaged in F. novicida for 4 days before being transformed into E. coli . Controls were isolated directly from E. coli . Representative replicates are shown. Three independent experiments were performed.
    Spei Restriction Enzymes, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 98 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    spei restriction enzymes - by Bioz Stars, 2022-07
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    OSN-specific GABAR B1 and D2R knockout mice, Related to Figure 2 . (A) Gene targeting at Drd2 locus. We obtained germline transmission from one ES clone, HEPD064_5_D11. The Drd2 locus in Drd2 tm1a , Drd2 fl (= Drd tm1c ) and Drd cKO . Drd2 +/ tm1a mice were crossed with Flp mice to obtain Drd2 +/ fl . ( C ) Gene targeting at Gabbr1 locus. We obtained germline transmission from one ES clone, EPD0730_1_G04. ( D ) The Gabbr1 locus in Gabbr1 tm1a , Gabbr1 fl (= Gabbr1 tm1c ), and Gabbr1 cKO . Gabbr1 +/ tm1a mice were crossed with Flp mice to obtain Gabbr1 +/ fl . ( E-G ) Southern blot analysis on Drd2 +/+ and Drd2 tm1a /+ ( E , Drd2 probe), Gabbr1 +/+ and Gabbr1 tm1a /+ ( F , Gabbr1 probe); Drd2 tm1a /+ and Gabbr1 tm1a /+ ( G , neo r probe). ( H ) Immunostaining of D2R in the OB of OSN-specific Drd2 mutant mice. ( I ) Immunostaining of GABAR B1 in the OB of OSN-specific Gabbr1 mutant mice. Location of 5’ and 3’ arms for gene targeting and 5’ and 3’ DNA probes for southern blotting are shown in ( A ) and ( C ). KpnI (K), SpeI (S), XhoI (X) and ApaI (A).

    Journal: bioRxiv

    Article Title: Widespread inhibition, antagonism, and synergy in mouse olfactory sensory neurons in vivo

    doi: 10.1101/803908

    Figure Lengend Snippet: OSN-specific GABAR B1 and D2R knockout mice, Related to Figure 2 . (A) Gene targeting at Drd2 locus. We obtained germline transmission from one ES clone, HEPD064_5_D11. The Drd2 locus in Drd2 tm1a , Drd2 fl (= Drd tm1c ) and Drd cKO . Drd2 +/ tm1a mice were crossed with Flp mice to obtain Drd2 +/ fl . ( C ) Gene targeting at Gabbr1 locus. We obtained germline transmission from one ES clone, EPD0730_1_G04. ( D ) The Gabbr1 locus in Gabbr1 tm1a , Gabbr1 fl (= Gabbr1 tm1c ), and Gabbr1 cKO . Gabbr1 +/ tm1a mice were crossed with Flp mice to obtain Gabbr1 +/ fl . ( E-G ) Southern blot analysis on Drd2 +/+ and Drd2 tm1a /+ ( E , Drd2 probe), Gabbr1 +/+ and Gabbr1 tm1a /+ ( F , Gabbr1 probe); Drd2 tm1a /+ and Gabbr1 tm1a /+ ( G , neo r probe). ( H ) Immunostaining of D2R in the OB of OSN-specific Drd2 mutant mice. ( I ) Immunostaining of GABAR B1 in the OB of OSN-specific Gabbr1 mutant mice. Location of 5’ and 3’ arms for gene targeting and 5’ and 3’ DNA probes for southern blotting are shown in ( A ) and ( C ). KpnI (K), SpeI (S), XhoI (X) and ApaI (A).

    Article Snippet: Southern blotting Genomic DNA (10 μg) extracted from the mouse tail was digested with restriction enzymes: KpnI (Toyobo), ApaI (NEB, R0114S), SpeI (NEB, R0133S), and XhoI (NEB, R0146S).

    Techniques: Knock-Out, Mouse Assay, Transmission Assay, Southern Blot, Immunostaining, Mutagenesis

    Plasmid stability. GFP expression was induced with 500 ng/ml of ATc. ON culture of day 0 was supplemented with 15 μg/ml kanamycin, while ON cultures of days 1–4 were supplemented with 50 μg/ml ampicillin. (A) GFP expression in F. novicida U112 pFNMB1 msfgfp ON cultures of day 0 and day 4. (B) GFP expression in F. novicida U112 pFNMB2 msfgfp ON cultures of day 0 and day 4. (A,B) Images are a merge of phase contrast and GFP channel. 26 × 39 μm fields of view are shown. Scale bar represent 5 μm. Representative replicates are shown. Three independent experiments were performed. (C) Survival assay performed with ON cultures of F. novicida U112 pFNMB1 msfgfp and F. novicida U112 pFNMB2 msfgfp plated on ampicillin and ampicillin/kanamycin plates. Three independent experiments were performed. (D) Digestion of pFNMB1 msfgfp with SacI and SpeI restriction enzymes. (E) Digestion of pFNMB2 msfgfp with SacI and SpeI restriction enzymes. (D,E) Plasmids were passaged in F. novicida for 4 days before being transformed into E. coli . Controls were isolated directly from E. coli . Representative replicates are shown. Three independent experiments were performed.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mobilizable Plasmids for Tunable Gene Expression in Francisella novicida

    doi: 10.3389/fcimb.2018.00284

    Figure Lengend Snippet: Plasmid stability. GFP expression was induced with 500 ng/ml of ATc. ON culture of day 0 was supplemented with 15 μg/ml kanamycin, while ON cultures of days 1–4 were supplemented with 50 μg/ml ampicillin. (A) GFP expression in F. novicida U112 pFNMB1 msfgfp ON cultures of day 0 and day 4. (B) GFP expression in F. novicida U112 pFNMB2 msfgfp ON cultures of day 0 and day 4. (A,B) Images are a merge of phase contrast and GFP channel. 26 × 39 μm fields of view are shown. Scale bar represent 5 μm. Representative replicates are shown. Three independent experiments were performed. (C) Survival assay performed with ON cultures of F. novicida U112 pFNMB1 msfgfp and F. novicida U112 pFNMB2 msfgfp plated on ampicillin and ampicillin/kanamycin plates. Three independent experiments were performed. (D) Digestion of pFNMB1 msfgfp with SacI and SpeI restriction enzymes. (E) Digestion of pFNMB2 msfgfp with SacI and SpeI restriction enzymes. (D,E) Plasmids were passaged in F. novicida for 4 days before being transformed into E. coli . Controls were isolated directly from E. coli . Representative replicates are shown. Three independent experiments were performed.

    Article Snippet: The plasmids were recovered again, digested with SacI and SpeI restriction enzymes and loaded on an agarose gel to analyze the size of the DNA fragments.

    Techniques: Plasmid Preparation, Expressing, Clonogenic Cell Survival Assay, Transformation Assay, Isolation

    PFGE patterns of SpeI-cleaved genomic DNA of 32 pure cultures obtained from END-49 . Assignment of the bacterial strains to Genome Group I, II, III, IV or V was indicated at the bottom of the PFGE photo.

    Journal: BMC Microbiology

    Article Title: Production of enterodiol from defatted flaxseeds through biotransformation by human intestinal bacteria

    doi: 10.1186/1471-2180-10-115

    Figure Lengend Snippet: PFGE patterns of SpeI-cleaved genomic DNA of 32 pure cultures obtained from END-49 . Assignment of the bacterial strains to Genome Group I, II, III, IV or V was indicated at the bottom of the PFGE photo.

    Article Snippet: Pulsed field gel electrophoresis (PFGE) The endonucleases I-CeuI, AvrII, XbaI and SpeI were purchased from New England Biolabs.

    Techniques:

    Plasmid stability. GFP expression was induced with 500 ng/ml of ATc. ON culture of day 0 was supplemented with 15 μg/ml kanamycin, while ON cultures of days 1–4 were supplemented with 50 μg/ml ampicillin. (A) GFP expression in F. novicida U112 pFNMB1 msfgfp ON cultures of day 0 and day 4. (B) GFP expression in F. novicida U112 pFNMB2 msfgfp ON cultures of day 0 and day 4. (A,B) Images are a merge of phase contrast and GFP channel. 26 × 39 μm fields of view are shown. Scale bar represent 5 μm. Representative replicates are shown. Three independent experiments were performed. (C) Survival assay performed with ON cultures of F. novicida U112 pFNMB1 msfgfp and F. novicida U112 pFNMB2 msfgfp plated on ampicillin and ampicillin/kanamycin plates. Three independent experiments were performed. (D) Digestion of pFNMB1 msfgfp with SacI and SpeI restriction enzymes. (E) Digestion of pFNMB2 msfgfp with SacI and SpeI restriction enzymes. (D,E) Plasmids were passaged in F. novicida for 4 days before being transformed into E. coli . Controls were isolated directly from E. coli . Representative replicates are shown. Three independent experiments were performed.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Mobilizable Plasmids for Tunable Gene Expression in Francisella novicida

    doi: 10.3389/fcimb.2018.00284

    Figure Lengend Snippet: Plasmid stability. GFP expression was induced with 500 ng/ml of ATc. ON culture of day 0 was supplemented with 15 μg/ml kanamycin, while ON cultures of days 1–4 were supplemented with 50 μg/ml ampicillin. (A) GFP expression in F. novicida U112 pFNMB1 msfgfp ON cultures of day 0 and day 4. (B) GFP expression in F. novicida U112 pFNMB2 msfgfp ON cultures of day 0 and day 4. (A,B) Images are a merge of phase contrast and GFP channel. 26 × 39 μm fields of view are shown. Scale bar represent 5 μm. Representative replicates are shown. Three independent experiments were performed. (C) Survival assay performed with ON cultures of F. novicida U112 pFNMB1 msfgfp and F. novicida U112 pFNMB2 msfgfp plated on ampicillin and ampicillin/kanamycin plates. Three independent experiments were performed. (D) Digestion of pFNMB1 msfgfp with SacI and SpeI restriction enzymes. (E) Digestion of pFNMB2 msfgfp with SacI and SpeI restriction enzymes. (D,E) Plasmids were passaged in F. novicida for 4 days before being transformed into E. coli . Controls were isolated directly from E. coli . Representative replicates are shown. Three independent experiments were performed.

    Article Snippet: The plasmids were recovered again, digested with SacI and SpeI restriction enzymes and loaded on an agarose gel to analyze the size of the DNA fragments.

    Techniques: Plasmid Preparation, Expressing, Clonogenic Cell Survival Assay, Transformation Assay, Isolation