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Stratagene long range pcr
Expression patterns of other neuronal innexins . (A) Predicted gene structures of neuronal innexins. Arrows indicate primer binding sites used for <t>PCR-amplified</t> green fluorescent protein <t>(GFP)</t> constructs (see Materials and methods). (B) INX-1::GFP expression (green) in motor neurons; DAPI stained nuclei in red. (C) INX-1::GFP expression (green) in dorsal nerve cord (dnc) and body wall muscles (bwm); dorsal nerve cord is visualized with anti-UNC-33 antibody (red). (D) INX-19::GFP expression in AVA and AVB interneurons. (E) INX-4::GFP expression in sensory neurons and pharyngeal m1 muscle cell. Abbreviations: nr, nerve ring.
Long Range Pcr, 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/long range pcr/product/Stratagene
Average 92 stars, based on 28 article reviews
Price from $9.99 to $1999.99
long range pcr - by Bioz Stars, 2021-02
92/100 stars

Images

1) Product Images from "Interactions between innexins UNC-7 and UNC-9 mediate electrical synapse specificity in the Caenorhabditis elegans locomotory nervous system"

Article Title: Interactions between innexins UNC-7 and UNC-9 mediate electrical synapse specificity in the Caenorhabditis elegans locomotory nervous system

Journal: Neural Development

doi: 10.1186/1749-8104-4-16

Expression patterns of other neuronal innexins . (A) Predicted gene structures of neuronal innexins. Arrows indicate primer binding sites used for PCR-amplified green fluorescent protein (GFP) constructs (see Materials and methods). (B) INX-1::GFP expression (green) in motor neurons; DAPI stained nuclei in red. (C) INX-1::GFP expression (green) in dorsal nerve cord (dnc) and body wall muscles (bwm); dorsal nerve cord is visualized with anti-UNC-33 antibody (red). (D) INX-19::GFP expression in AVA and AVB interneurons. (E) INX-4::GFP expression in sensory neurons and pharyngeal m1 muscle cell. Abbreviations: nr, nerve ring.
Figure Legend Snippet: Expression patterns of other neuronal innexins . (A) Predicted gene structures of neuronal innexins. Arrows indicate primer binding sites used for PCR-amplified green fluorescent protein (GFP) constructs (see Materials and methods). (B) INX-1::GFP expression (green) in motor neurons; DAPI stained nuclei in red. (C) INX-1::GFP expression (green) in dorsal nerve cord (dnc) and body wall muscles (bwm); dorsal nerve cord is visualized with anti-UNC-33 antibody (red). (D) INX-19::GFP expression in AVA and AVB interneurons. (E) INX-4::GFP expression in sensory neurons and pharyngeal m1 muscle cell. Abbreviations: nr, nerve ring.

Techniques Used: Expressing, Binding Assay, Polymerase Chain Reaction, Amplification, Construct, Staining, Antiviral Assay

2) Product Images from "laminin alpha 1 gene is essential for normal lens development in zebrafish"

Article Title: laminin alpha 1 gene is essential for normal lens development in zebrafish

Journal: BMC Developmental Biology

doi: 10.1186/1471-213X-6-13

Expression of zebrafish laminin alpha 1 gene . I. RT-PCR analysis of lama1 expression in embryos and adult fish . RT-PCR results for lama1, lama2 and control bactin transcripts are presented as indicated. Embryonic (16-32 cells to 120-hpf) or adult (1 year old) cDNA samples employed in reactions are indicated at the top: lane 1- 16-32 cells, 2- 3-8 hpf, 3- 24 hpf, 4- 36 hpf, 5- 48 hpf, 6- 72 hpf, 7- 84 hpf, 8- 120 hpf embryos; for adult tissues- lane 9 contains products obtained with adult eye cDNA, 10- brain, 11- jaws, 12- internal organs and 13- tail. II . In situ hybridization of antisense lama1 riboprobe in zebrafish embryos . A-F : 8-96 hpf whole zebrafish embryos that were hybridized with lama1 DIG-labeled antisense riboprobe. G-L : Transverse sections of 48-96 hpf zebrafish embryos at the level of the eye ( G ), brain ( H ), otic vesicle ( I ), developing kidney ( J ), and trunk ( K , L ). Embryonic stages are indicated at the bottom of the picture. At 8-hpf, expression of the lama1 gene was detected in all embryonic tissues; by 24-hpf, higher levels of transcript were evident in the developing lens (arrows in B-E; le in F and G) and sclera ( sc ) of the eye, brain ( b ), somites ( s ), and otic vesicle ( ov ), pronephros ( p ) and pronephric duct ( pd ), notochord ( n ). e - eye, m -midbrain.
Figure Legend Snippet: Expression of zebrafish laminin alpha 1 gene . I. RT-PCR analysis of lama1 expression in embryos and adult fish . RT-PCR results for lama1, lama2 and control bactin transcripts are presented as indicated. Embryonic (16-32 cells to 120-hpf) or adult (1 year old) cDNA samples employed in reactions are indicated at the top: lane 1- 16-32 cells, 2- 3-8 hpf, 3- 24 hpf, 4- 36 hpf, 5- 48 hpf, 6- 72 hpf, 7- 84 hpf, 8- 120 hpf embryos; for adult tissues- lane 9 contains products obtained with adult eye cDNA, 10- brain, 11- jaws, 12- internal organs and 13- tail. II . In situ hybridization of antisense lama1 riboprobe in zebrafish embryos . A-F : 8-96 hpf whole zebrafish embryos that were hybridized with lama1 DIG-labeled antisense riboprobe. G-L : Transverse sections of 48-96 hpf zebrafish embryos at the level of the eye ( G ), brain ( H ), otic vesicle ( I ), developing kidney ( J ), and trunk ( K , L ). Embryonic stages are indicated at the bottom of the picture. At 8-hpf, expression of the lama1 gene was detected in all embryonic tissues; by 24-hpf, higher levels of transcript were evident in the developing lens (arrows in B-E; le in F and G) and sclera ( sc ) of the eye, brain ( b ), somites ( s ), and otic vesicle ( ov ), pronephros ( p ) and pronephric duct ( pd ), notochord ( n ). e - eye, m -midbrain.

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction, Fluorescence In Situ Hybridization, In Situ Hybridization, Labeling

3) Product Images from "Hemolysin of Uropathogenic Escherichia coli Evokes Extensive Shedding of the Uroepithelium and Hemorrhage in Bladder Tissue within the First 24 Hours after Intraurethral Inoculation of Mice "

Article Title: Hemolysin of Uropathogenic Escherichia coli Evokes Extensive Shedding of the Uroepithelium and Hemorrhage in Bladder Tissue within the First 24 Hours after Intraurethral Inoculation of Mice

Journal: Infection and Immunity

doi: 10.1128/IAI.00075-08

Long-range PCR. (A) Primers KG09 and KG10R (arrows above the hemolysin operon) were used to test for the presence of an hlyA 1 -containing PCR fragment amplified from hlyA 1 to the cnf 1 gene on the chromosome of the following strains: CP9 (lane 1), CP9 cnf 1 (lane 2), CP9Δ hlyA 1 :: cat (lane 3), and CP9 cnf 1 Δ hlyA 1 :: cat (lane 4). PCR products (∼6.7 kb) were present for CP9 and CP9 cnf 1 but not for CP9Δ hlyA 1 :: cat and CP9 cnf 1 Δ hlyA 1 :: cat , as shown in the top panel of the ethidium bromide-stained agarose gel on the left. Primers KG03F and KG10R were used to verify the replacement of hlyA 1 with cat upstream of the cnf 1 gene in mutant strains CP9Δ hlyA 1 :: cat and CP9 cnf 1 Δ hlyA 1 :: cat (∼5.0-kb amplified PCR product from cat to cnf 1 in the bottom panel of the ethidium bromide-stained agarose gel). (B) Western blot analysis with polyclonal anti-CNF1 serum as a probe was used to verify that wild-type levels of CNF1 (lane 1) were produced by CP9Δ hlyA 1 :: cat (lane 3) and that, as expected, strains CP9 cnf 1 and CP9 cnf 1 Δ hlyA 1 :: cat did not produce CNF1 (lanes 2 and 4, respectively).
Figure Legend Snippet: Long-range PCR. (A) Primers KG09 and KG10R (arrows above the hemolysin operon) were used to test for the presence of an hlyA 1 -containing PCR fragment amplified from hlyA 1 to the cnf 1 gene on the chromosome of the following strains: CP9 (lane 1), CP9 cnf 1 (lane 2), CP9Δ hlyA 1 :: cat (lane 3), and CP9 cnf 1 Δ hlyA 1 :: cat (lane 4). PCR products (∼6.7 kb) were present for CP9 and CP9 cnf 1 but not for CP9Δ hlyA 1 :: cat and CP9 cnf 1 Δ hlyA 1 :: cat , as shown in the top panel of the ethidium bromide-stained agarose gel on the left. Primers KG03F and KG10R were used to verify the replacement of hlyA 1 with cat upstream of the cnf 1 gene in mutant strains CP9Δ hlyA 1 :: cat and CP9 cnf 1 Δ hlyA 1 :: cat (∼5.0-kb amplified PCR product from cat to cnf 1 in the bottom panel of the ethidium bromide-stained agarose gel). (B) Western blot analysis with polyclonal anti-CNF1 serum as a probe was used to verify that wild-type levels of CNF1 (lane 1) were produced by CP9Δ hlyA 1 :: cat (lane 3) and that, as expected, strains CP9 cnf 1 and CP9 cnf 1 Δ hlyA 1 :: cat did not produce CNF1 (lanes 2 and 4, respectively).

Techniques Used: Polymerase Chain Reaction, Amplification, Staining, Agarose Gel Electrophoresis, Mutagenesis, Western Blot, Produced

Related Articles

Clone Assay:

Article Title: CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells
Article Snippet: .. The left and right arms of the disruption construct were amplified as 2.0 and 3.5 kb fragments by long-range PCR with a genomic clone as the template, and were cloned into pBluescript (Stratagene). .. For the CENP-H-puro disruption construct, the puro-resistance cassette under control of the β-actin promoter was inserted between the two arms.

Article Title: laminin alpha 1 gene is essential for normal lens development in zebrafish
Article Snippet: .. Cloning of lama1: RT-PCR, RACE, long-range PCR, cloning and sequencing PCR products were generated using specific oligos, PfuUltra high-fidelity DNA polymerase (Stratagene, La Jolla, CA) and standard conditions described elsewhere [ ]. .. The PCR products were separated by electrophoresis in 1% agarose gel, cloned into a pCRII-TOPO vector (Invitrogen, Carlsbad, CA) and subjected to DNA sequencing using the ABI PRISM 373 DNA Sequencer.

Amplification:

Article Title: CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells
Article Snippet: .. The left and right arms of the disruption construct were amplified as 2.0 and 3.5 kb fragments by long-range PCR with a genomic clone as the template, and were cloned into pBluescript (Stratagene). .. For the CENP-H-puro disruption construct, the puro-resistance cassette under control of the β-actin promoter was inserted between the two arms.

Polymerase Chain Reaction:

Article Title: CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells
Article Snippet: .. The left and right arms of the disruption construct were amplified as 2.0 and 3.5 kb fragments by long-range PCR with a genomic clone as the template, and were cloned into pBluescript (Stratagene). .. For the CENP-H-puro disruption construct, the puro-resistance cassette under control of the β-actin promoter was inserted between the two arms.

Article Title: An 85-kb tandem triplication in the slow Wallerian degeneration (Wlds) mouse
Article Snippet: .. The method of Saiki et al. ( ) was followed by using Taq DNA polymerase (Promega) or, for long-range PCR or products for sequencing, Pfu polymerase (Stratagene). .. Custom oligonucleotides were obtained from GIBCO/BRL and D4Mit map pairs from Research Genetics.

Article Title: Unc-45 Mutations in Caenorhabditis elegans Implicate a CRO1/She4p-like Domain in Myosin Assembly
Article Snippet: .. Long range PCR, using Taq Extender PCR Additive (Stratagene, La Jolla, CA) and Taq DNA Polymerase ( Promega Corp. , Madison, WI), was performed with an aliquot of the Okemma library as template. ..

Article Title: laminin alpha 1 gene is essential for normal lens development in zebrafish
Article Snippet: .. Cloning of lama1: RT-PCR, RACE, long-range PCR, cloning and sequencing PCR products were generated using specific oligos, PfuUltra high-fidelity DNA polymerase (Stratagene, La Jolla, CA) and standard conditions described elsewhere [ ]. .. The PCR products were separated by electrophoresis in 1% agarose gel, cloned into a pCRII-TOPO vector (Invitrogen, Carlsbad, CA) and subjected to DNA sequencing using the ABI PRISM 373 DNA Sequencer.

Article Title: Neurotransmitter Transporter-Like: A Male Germline-specific SLC6 Transporter Required for Drosophila Spermiogenesis
Article Snippet: .. Long Range PCR was done on the repaired clone (AT20383) using PfuTurbo DNA Polymerase (Stratagene) to attach Not I sites at either end for ligating into pTMR vector (a gift from Dr. Ming Guo). ..

Article Title: Hemolysin of Uropathogenic Escherichia coli Evokes Extensive Shedding of the Uroepithelium and Hemorrhage in Bladder Tissue within the First 24 Hours after Intraurethral Inoculation of Mice
Article Snippet: .. The location of the hlyA 1 mutation in the hly operon adjacent to the cnf 1 locus was confirmed by performing long-range PCR with Pfu TURBO (Stratagene, La Jolla, CA) and the following primer sets: KG03F-KG10R and KG09F-KG10R (Fig. ). .. The CP9 cnf 1 Δ hlyA 1 :: cat mutant grew at wild-type levels and exhibited a reduced hemolytic phenotype on blood agar plates compared to CP9 (not shown).

Article Title: Interactions between innexins UNC-7 and UNC-9 mediate electrical synapse specificity in the Caenorhabditis elegans locomotory nervous system
Article Snippet: .. unc-9 rescue and innexin::gfp constructions using long-range PCR A plasmid encompassing the unc-9 gene from exon 4 to the polyadenylation site at nucleotide 12,661 of cosmid R12H7 [ ] was constructed in pBluescript (Stratagene, Cedar Creek, TX, USA) extending from the Kpn I site at nucleotides 8,959–8,964 to the Xba I site at nucleotides 14,519–14,524. .. A PCR product from nucleotide 4,178 of F09D5 (primer UNC-9A: 5'-TCAGACGCGATGTTTTGTAGGTGGG-3') to nucleotide 9,849 of cosmid R12H7 (Primer UNC-9B: 5'-CCACTTGATGTTTCACGTGTAGCGC-3') was amplified (Expand Long Template PCR System, Roche Diagnostics, Indianapolis, IN, USA) from genomic DNA (14,678 nucleotides total length).

Mutagenesis:

Article Title: Hemolysin of Uropathogenic Escherichia coli Evokes Extensive Shedding of the Uroepithelium and Hemorrhage in Bladder Tissue within the First 24 Hours after Intraurethral Inoculation of Mice
Article Snippet: .. The location of the hlyA 1 mutation in the hly operon adjacent to the cnf 1 locus was confirmed by performing long-range PCR with Pfu TURBO (Stratagene, La Jolla, CA) and the following primer sets: KG03F-KG10R and KG09F-KG10R (Fig. ). .. The CP9 cnf 1 Δ hlyA 1 :: cat mutant grew at wild-type levels and exhibited a reduced hemolytic phenotype on blood agar plates compared to CP9 (not shown).

Subcloning:

Article Title: The ACA4 Gene of Arabidopsis Encodes a Vacuolar Membrane Calcium Pump That Improves Salt Tolerance in Yeast 1
Article Snippet: .. For sub-cloning of the entire gene, a long-range nested PCR on cDNA derived from library CD4-16 was performed as described above using the Taq + Precision system (Stratagene). .. The nested primers were designed to contain restriction sites for directional cloning; additionally, the primer matching the 5′ end contained a sequence coding for a C-terminal RGSH6 motif enabling western detection and purification by Ni2+ -affinity chromatography.

Construct:

Article Title: CENP-H, a constitutive centromere component, is required for centromere targeting of CENP-C in vertebrate cells
Article Snippet: .. The left and right arms of the disruption construct were amplified as 2.0 and 3.5 kb fragments by long-range PCR with a genomic clone as the template, and were cloned into pBluescript (Stratagene). .. For the CENP-H-puro disruption construct, the puro-resistance cassette under control of the β-actin promoter was inserted between the two arms.

Article Title: Interactions between innexins UNC-7 and UNC-9 mediate electrical synapse specificity in the Caenorhabditis elegans locomotory nervous system
Article Snippet: .. unc-9 rescue and innexin::gfp constructions using long-range PCR A plasmid encompassing the unc-9 gene from exon 4 to the polyadenylation site at nucleotide 12,661 of cosmid R12H7 [ ] was constructed in pBluescript (Stratagene, Cedar Creek, TX, USA) extending from the Kpn I site at nucleotides 8,959–8,964 to the Xba I site at nucleotides 14,519–14,524. .. A PCR product from nucleotide 4,178 of F09D5 (primer UNC-9A: 5'-TCAGACGCGATGTTTTGTAGGTGGG-3') to nucleotide 9,849 of cosmid R12H7 (Primer UNC-9B: 5'-CCACTTGATGTTTCACGTGTAGCGC-3') was amplified (Expand Long Template PCR System, Roche Diagnostics, Indianapolis, IN, USA) from genomic DNA (14,678 nucleotides total length).

Sequencing:

Article Title: An 85-kb tandem triplication in the slow Wallerian degeneration (Wlds) mouse
Article Snippet: .. The method of Saiki et al. ( ) was followed by using Taq DNA polymerase (Promega) or, for long-range PCR or products for sequencing, Pfu polymerase (Stratagene). .. Custom oligonucleotides were obtained from GIBCO/BRL and D4Mit map pairs from Research Genetics.

Article Title: laminin alpha 1 gene is essential for normal lens development in zebrafish
Article Snippet: .. Cloning of lama1: RT-PCR, RACE, long-range PCR, cloning and sequencing PCR products were generated using specific oligos, PfuUltra high-fidelity DNA polymerase (Stratagene, La Jolla, CA) and standard conditions described elsewhere [ ]. .. The PCR products were separated by electrophoresis in 1% agarose gel, cloned into a pCRII-TOPO vector (Invitrogen, Carlsbad, CA) and subjected to DNA sequencing using the ABI PRISM 373 DNA Sequencer.

Generated:

Article Title: laminin alpha 1 gene is essential for normal lens development in zebrafish
Article Snippet: .. Cloning of lama1: RT-PCR, RACE, long-range PCR, cloning and sequencing PCR products were generated using specific oligos, PfuUltra high-fidelity DNA polymerase (Stratagene, La Jolla, CA) and standard conditions described elsewhere [ ]. .. The PCR products were separated by electrophoresis in 1% agarose gel, cloned into a pCRII-TOPO vector (Invitrogen, Carlsbad, CA) and subjected to DNA sequencing using the ABI PRISM 373 DNA Sequencer.

Plasmid Preparation:

Article Title: Neurotransmitter Transporter-Like: A Male Germline-specific SLC6 Transporter Required for Drosophila Spermiogenesis
Article Snippet: .. Long Range PCR was done on the repaired clone (AT20383) using PfuTurbo DNA Polymerase (Stratagene) to attach Not I sites at either end for ligating into pTMR vector (a gift from Dr. Ming Guo). ..

Article Title: Interactions between innexins UNC-7 and UNC-9 mediate electrical synapse specificity in the Caenorhabditis elegans locomotory nervous system
Article Snippet: .. unc-9 rescue and innexin::gfp constructions using long-range PCR A plasmid encompassing the unc-9 gene from exon 4 to the polyadenylation site at nucleotide 12,661 of cosmid R12H7 [ ] was constructed in pBluescript (Stratagene, Cedar Creek, TX, USA) extending from the Kpn I site at nucleotides 8,959–8,964 to the Xba I site at nucleotides 14,519–14,524. .. A PCR product from nucleotide 4,178 of F09D5 (primer UNC-9A: 5'-TCAGACGCGATGTTTTGTAGGTGGG-3') to nucleotide 9,849 of cosmid R12H7 (Primer UNC-9B: 5'-CCACTTGATGTTTCACGTGTAGCGC-3') was amplified (Expand Long Template PCR System, Roche Diagnostics, Indianapolis, IN, USA) from genomic DNA (14,678 nucleotides total length).

Reverse Transcription Polymerase Chain Reaction:

Article Title: laminin alpha 1 gene is essential for normal lens development in zebrafish
Article Snippet: .. Cloning of lama1: RT-PCR, RACE, long-range PCR, cloning and sequencing PCR products were generated using specific oligos, PfuUltra high-fidelity DNA polymerase (Stratagene, La Jolla, CA) and standard conditions described elsewhere [ ]. .. The PCR products were separated by electrophoresis in 1% agarose gel, cloned into a pCRII-TOPO vector (Invitrogen, Carlsbad, CA) and subjected to DNA sequencing using the ABI PRISM 373 DNA Sequencer.

Nested PCR:

Article Title: The ACA4 Gene of Arabidopsis Encodes a Vacuolar Membrane Calcium Pump That Improves Salt Tolerance in Yeast 1
Article Snippet: .. For sub-cloning of the entire gene, a long-range nested PCR on cDNA derived from library CD4-16 was performed as described above using the Taq + Precision system (Stratagene). .. The nested primers were designed to contain restriction sites for directional cloning; additionally, the primer matching the 5′ end contained a sequence coding for a C-terminal RGSH6 motif enabling western detection and purification by Ni2+ -affinity chromatography.

Derivative Assay:

Article Title: The ACA4 Gene of Arabidopsis Encodes a Vacuolar Membrane Calcium Pump That Improves Salt Tolerance in Yeast 1
Article Snippet: .. For sub-cloning of the entire gene, a long-range nested PCR on cDNA derived from library CD4-16 was performed as described above using the Taq + Precision system (Stratagene). .. The nested primers were designed to contain restriction sites for directional cloning; additionally, the primer matching the 5′ end contained a sequence coding for a C-terminal RGSH6 motif enabling western detection and purification by Ni2+ -affinity chromatography.

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    Stratagene long range pcr
    Expression patterns of other neuronal innexins . (A) Predicted gene structures of neuronal innexins. Arrows indicate primer binding sites used for <t>PCR-amplified</t> green fluorescent protein <t>(GFP)</t> constructs (see Materials and methods). (B) INX-1::GFP expression (green) in motor neurons; DAPI stained nuclei in red. (C) INX-1::GFP expression (green) in dorsal nerve cord (dnc) and body wall muscles (bwm); dorsal nerve cord is visualized with anti-UNC-33 antibody (red). (D) INX-19::GFP expression in AVA and AVB interneurons. (E) INX-4::GFP expression in sensory neurons and pharyngeal m1 muscle cell. Abbreviations: nr, nerve ring.
    Long Range Pcr, supplied by Stratagene, used in various techniques. Bioz Stars score: 92/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/long range pcr/product/Stratagene
    Average 92 stars, based on 30 article reviews
    Price from $9.99 to $1999.99
    long range pcr - by Bioz Stars, 2021-02
    92/100 stars
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    Expression patterns of other neuronal innexins . (A) Predicted gene structures of neuronal innexins. Arrows indicate primer binding sites used for PCR-amplified green fluorescent protein (GFP) constructs (see Materials and methods). (B) INX-1::GFP expression (green) in motor neurons; DAPI stained nuclei in red. (C) INX-1::GFP expression (green) in dorsal nerve cord (dnc) and body wall muscles (bwm); dorsal nerve cord is visualized with anti-UNC-33 antibody (red). (D) INX-19::GFP expression in AVA and AVB interneurons. (E) INX-4::GFP expression in sensory neurons and pharyngeal m1 muscle cell. Abbreviations: nr, nerve ring.

    Journal: Neural Development

    Article Title: Interactions between innexins UNC-7 and UNC-9 mediate electrical synapse specificity in the Caenorhabditis elegans locomotory nervous system

    doi: 10.1186/1749-8104-4-16

    Figure Lengend Snippet: Expression patterns of other neuronal innexins . (A) Predicted gene structures of neuronal innexins. Arrows indicate primer binding sites used for PCR-amplified green fluorescent protein (GFP) constructs (see Materials and methods). (B) INX-1::GFP expression (green) in motor neurons; DAPI stained nuclei in red. (C) INX-1::GFP expression (green) in dorsal nerve cord (dnc) and body wall muscles (bwm); dorsal nerve cord is visualized with anti-UNC-33 antibody (red). (D) INX-19::GFP expression in AVA and AVB interneurons. (E) INX-4::GFP expression in sensory neurons and pharyngeal m1 muscle cell. Abbreviations: nr, nerve ring.

    Article Snippet: unc-9 rescue and innexin::gfp constructions using long-range PCR A plasmid encompassing the unc-9 gene from exon 4 to the polyadenylation site at nucleotide 12,661 of cosmid R12H7 [ ] was constructed in pBluescript (Stratagene, Cedar Creek, TX, USA) extending from the Kpn I site at nucleotides 8,959–8,964 to the Xba I site at nucleotides 14,519–14,524.

    Techniques: Expressing, Binding Assay, Polymerase Chain Reaction, Amplification, Construct, Staining, Antiviral Assay

    ACA4 expression is throughout the plant and is induced by salt stress. A, ACA4-specific transcripts (ACA4) were monitored in total RNA from different tissues by RT-PCR amplification of a 561-bp cDNA fragment. To circumvent problems with genomic DNA contamination, primers for ACA4 were chosen to cover an intron-spanning region (intron 3). As an internal control, RT-PCR on the actin Aac1 gene was employed. Control RT-PCR amplification yielded essentially constant signals (

    Journal: Plant Physiology

    Article Title: The ACA4 Gene of Arabidopsis Encodes a Vacuolar Membrane Calcium Pump That Improves Salt Tolerance in Yeast 1

    doi:

    Figure Lengend Snippet: ACA4 expression is throughout the plant and is induced by salt stress. A, ACA4-specific transcripts (ACA4) were monitored in total RNA from different tissues by RT-PCR amplification of a 561-bp cDNA fragment. To circumvent problems with genomic DNA contamination, primers for ACA4 were chosen to cover an intron-spanning region (intron 3). As an internal control, RT-PCR on the actin Aac1 gene was employed. Control RT-PCR amplification yielded essentially constant signals (

    Article Snippet: For sub-cloning of the entire gene, a long-range nested PCR on cDNA derived from library CD4-16 was performed as described above using the Taq + Precision system (Stratagene).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Amplification

    Expression of zebrafish laminin alpha 1 gene . I. RT-PCR analysis of lama1 expression in embryos and adult fish . RT-PCR results for lama1, lama2 and control bactin transcripts are presented as indicated. Embryonic (16-32 cells to 120-hpf) or adult (1 year old) cDNA samples employed in reactions are indicated at the top: lane 1- 16-32 cells, 2- 3-8 hpf, 3- 24 hpf, 4- 36 hpf, 5- 48 hpf, 6- 72 hpf, 7- 84 hpf, 8- 120 hpf embryos; for adult tissues- lane 9 contains products obtained with adult eye cDNA, 10- brain, 11- jaws, 12- internal organs and 13- tail. II . In situ hybridization of antisense lama1 riboprobe in zebrafish embryos . A-F : 8-96 hpf whole zebrafish embryos that were hybridized with lama1 DIG-labeled antisense riboprobe. G-L : Transverse sections of 48-96 hpf zebrafish embryos at the level of the eye ( G ), brain ( H ), otic vesicle ( I ), developing kidney ( J ), and trunk ( K , L ). Embryonic stages are indicated at the bottom of the picture. At 8-hpf, expression of the lama1 gene was detected in all embryonic tissues; by 24-hpf, higher levels of transcript were evident in the developing lens (arrows in B-E; le in F and G) and sclera ( sc ) of the eye, brain ( b ), somites ( s ), and otic vesicle ( ov ), pronephros ( p ) and pronephric duct ( pd ), notochord ( n ). e - eye, m -midbrain.

    Journal: BMC Developmental Biology

    Article Title: laminin alpha 1 gene is essential for normal lens development in zebrafish

    doi: 10.1186/1471-213X-6-13

    Figure Lengend Snippet: Expression of zebrafish laminin alpha 1 gene . I. RT-PCR analysis of lama1 expression in embryos and adult fish . RT-PCR results for lama1, lama2 and control bactin transcripts are presented as indicated. Embryonic (16-32 cells to 120-hpf) or adult (1 year old) cDNA samples employed in reactions are indicated at the top: lane 1- 16-32 cells, 2- 3-8 hpf, 3- 24 hpf, 4- 36 hpf, 5- 48 hpf, 6- 72 hpf, 7- 84 hpf, 8- 120 hpf embryos; for adult tissues- lane 9 contains products obtained with adult eye cDNA, 10- brain, 11- jaws, 12- internal organs and 13- tail. II . In situ hybridization of antisense lama1 riboprobe in zebrafish embryos . A-F : 8-96 hpf whole zebrafish embryos that were hybridized with lama1 DIG-labeled antisense riboprobe. G-L : Transverse sections of 48-96 hpf zebrafish embryos at the level of the eye ( G ), brain ( H ), otic vesicle ( I ), developing kidney ( J ), and trunk ( K , L ). Embryonic stages are indicated at the bottom of the picture. At 8-hpf, expression of the lama1 gene was detected in all embryonic tissues; by 24-hpf, higher levels of transcript were evident in the developing lens (arrows in B-E; le in F and G) and sclera ( sc ) of the eye, brain ( b ), somites ( s ), and otic vesicle ( ov ), pronephros ( p ) and pronephric duct ( pd ), notochord ( n ). e - eye, m -midbrain.

    Article Snippet: Cloning of lama1: RT-PCR, RACE, long-range PCR, cloning and sequencing PCR products were generated using specific oligos, PfuUltra high-fidelity DNA polymerase (Stratagene, La Jolla, CA) and standard conditions described elsewhere [ ].

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Fluorescence In Situ Hybridization, In Situ Hybridization, Labeling

    Creation of Nod2 −/− mice. (a) Diagram showing the targeting strategy to delete the first exon of Nod2 encoding the first CARD domain and the start codon in addition to part of the second CARD domain. (b) Southern analysis of tail DNA isolated from a litter produced by a Nod2 +/− mouse cross-probed with the external probe shown in panel a. (c) RT-PCR analysis of cells isolated from Nod2 −/− mice. Total RNA was isolated from BMDMs, T or B cells, or CD11c+ enriched DCs from +/+ or Nod2 −/− mice. RNA was reverse transcribed and subjected to PCR analysis for Nod2 exon 1/2, Nod1, or actin (control). A diagram of the RT-PCR strategy for Nod2 is shown in panel d. A similar strategy to cross the first intron was adopted for Nod1.

    Journal: Molecular and Cellular Biology

    Article Title: Role of Nod2 in the Response of Macrophages to Toll-Like Receptor Agonists

    doi: 10.1128/MCB.23.21.7531-7539.2003

    Figure Lengend Snippet: Creation of Nod2 −/− mice. (a) Diagram showing the targeting strategy to delete the first exon of Nod2 encoding the first CARD domain and the start codon in addition to part of the second CARD domain. (b) Southern analysis of tail DNA isolated from a litter produced by a Nod2 +/− mouse cross-probed with the external probe shown in panel a. (c) RT-PCR analysis of cells isolated from Nod2 −/− mice. Total RNA was isolated from BMDMs, T or B cells, or CD11c+ enriched DCs from +/+ or Nod2 −/− mice. RNA was reverse transcribed and subjected to PCR analysis for Nod2 exon 1/2, Nod1, or actin (control). A diagram of the RT-PCR strategy for Nod2 is shown in panel d. A similar strategy to cross the first intron was adopted for Nod1.

    Article Snippet: The arms were isolated by high-fidelity long-range PCR (Herculase; Stratagene) from C57BL/6 genomic DNA.

    Techniques: Mouse Assay, Isolation, Produced, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction