expand long template pcr system  (Roche)


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    Structured Review

    Roche expand long template pcr system
    Expand Long Template Pcr System, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 1004 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/expand long template pcr system/product/Roche
    Average 92 stars, based on 1004 article reviews
    Price from $9.99 to $1999.99
    expand long template pcr system - by Bioz Stars, 2021-01
    92/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: The Mitochondrial Genome of Conus textile, coxI-coxII Intergenic Sequences and Conoidean Evolution
    Article Snippet: .. The Expand Long Template PCR System from Roche Molecular Biochemicals (Roche Diagnostics Corporation, Indianapolis,IN, USA) was used for these experiments. ..

    Article Title: A shared enhancer controls a temporal switch between promoters during Drosophila primary sex determination
    Article Snippet: .. Sxl genomic fragments were made using the Expand Long-Template polymerase chain reaction system (Roche), cloned into pCRII-TOPO (Invitrogen), and ligated into P-element transformation vectors based on pCaSpeR-AUG-ßgal. .. Germline transformations were performed by Genetic Services, Inc, Cambridge, MA.

    Article Title: Signal recognition particle (SRP)-mediated targeting and Sec-dependent translocation of an extracellular Escherichia coli protein.
    Article Snippet: .. Reagents and Sera—Restriction enzymes, the Expand long template PCR system and the Lumi-light Western blotting substrate were obtained from Roche Molecular Biochemicals. .. Antiserum J40 raised against purified Hbp has been described previously (2).

    Article Title: A Functional Genetic Polymorphism on Human Carbonyl Reductase 1 (CBR1 V88I) Impacts on Catalytic Activity and NADPH Binding Affinity
    Article Snippet: .. One microliter of the cDNA conversion mixture was amplified by using the Expand Long Template PCR system with buffer 1 (Roche Diagnostics, Indianapolis, IN). .. PCR amplification conditions were 95°C for 2 min; 45 cycles of 95°C for 35 s, 57.5°C for 40 s, and 68°C for 70 s; and final extension at 70°C for 10 min. Aliquots of PCR products were visualized by UV light after size-fractionation in 1.0% agarose gels supplemented with ethidium bromide (0.5 μg/ml).

    Article Title: Fibroblast growth factor 21 and grow differentiation factor 15 are sensitive biomarkers of mitochondrial diseases due to mitochondrial transfer-RNA mutations and mitochondrial DNA deletions.
    Article Snippet: .. Diagnosis of mitochondrial diseases (MDs) is challenging, since they are multisystemic disorders, characterized by a heterogeneous symptomatology.. Recently, an increase in serum levels of fibroblast growth factor 21 (FGF21) and growth differentiation factor 15 (GDF15) has been found in the majority of patients with MDs compared with healthy controls. ..

    Western Blot:

    Article Title: Signal recognition particle (SRP)-mediated targeting and Sec-dependent translocation of an extracellular Escherichia coli protein.
    Article Snippet: .. Reagents and Sera—Restriction enzymes, the Expand long template PCR system and the Lumi-light Western blotting substrate were obtained from Roche Molecular Biochemicals. .. Antiserum J40 raised against purified Hbp has been described previously (2).

    Transformation Assay:

    Article Title: A shared enhancer controls a temporal switch between promoters during Drosophila primary sex determination
    Article Snippet: .. Sxl genomic fragments were made using the Expand Long-Template polymerase chain reaction system (Roche), cloned into pCRII-TOPO (Invitrogen), and ligated into P-element transformation vectors based on pCaSpeR-AUG-ßgal. .. Germline transformations were performed by Genetic Services, Inc, Cambridge, MA.

    Amplification:

    Article Title: A Functional Genetic Polymorphism on Human Carbonyl Reductase 1 (CBR1 V88I) Impacts on Catalytic Activity and NADPH Binding Affinity
    Article Snippet: .. One microliter of the cDNA conversion mixture was amplified by using the Expand Long Template PCR system with buffer 1 (Roche Diagnostics, Indianapolis, IN). .. PCR amplification conditions were 95°C for 2 min; 45 cycles of 95°C for 35 s, 57.5°C for 40 s, and 68°C for 70 s; and final extension at 70°C for 10 min. Aliquots of PCR products were visualized by UV light after size-fractionation in 1.0% agarose gels supplemented with ethidium bromide (0.5 μg/ml).

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    Roche long range pcr
    Confirmation and characterization of the rearrangements . (A) Confirmation of the deletion of exons 1A/1B-2 by long-range <t>PCR</t> and sequencing of the breakpoints. (B) Confirmation of the deletion of exons 5–14 by long-range PCR and sequencing of the breakpoints. (C) Confirmation of the deletion of exons 11–12 by long-range PCR and sequencing of the breakpoints. Lanes 1+, 2+, carriers of the deletion; lane C-, negative control (wt); lane B, blank; lane M, marker (Ready-Load™ 1 Kb <t>DNA</t> Ladder, Invitrogen).
    Long Range Pcr, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 167 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/long range pcr/product/Roche
    Average 92 stars, based on 167 article reviews
    Price from $9.99 to $1999.99
    long range pcr - by Bioz Stars, 2021-01
    92/100 stars
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    92
    Roche expand long template pcr system
    Deletion of SAMHD1 does not affect <t>mtDNA</t> stability. a) MtDNA copy number in the TA muscle of 5 or 6 wt (filled dots) and SAMHD1 −/− (open dots) 13-week-old (adult), 1-year-old (old adult), and 2-year-old (aged) animals was determined by qPCR and normalized to the value for adult wt mice. The mean for each group is indicated by a horizontal line. The p-values were calculated using Welch’s t-test; ns, non-significant. b) DNA isolated from embryos and from the TA muscle of pups, adults, 1-year-old (old) adults, or aged animals was linearized with SacI endonuclease and separated on a neutral gel. MtDNA was visualized as above. Full-length mtDNA is indicated (FL); the asterisk denotes a higher-migrating species resistant to cleavage. c) Long-range <t>PCR</t> to detect deletions in mtDNA from the TA muscle of wt mice of various ages. Full-length product is indicated (FL). Only minor species containing deletions are observed in the mtDNA from old adults and aged animals, as indicated by the vertical line on the right-hand side of the gel. d) Untreated or alkali-treated DNA from skeletal muscle of aged wt and SAMHD1 −/− (ko) mice was analyzed on a denaturing gel, and mtDNA was visualized using a COX1 probe. Each sample lane corresponds to an individual mouse, and dotted lines represent the median. e) The median length of the untreated mtDNA in samples from Fig. 5d is indicated by a horizontal line. The two groups were compared using Welch’s t-test (ns, non-significant; n = 4). f) The length difference between untreated and alkali-treated mtDNAs shown in Fig. 5d was used to compute the number of rNMPs per single strand of mtDNA. The horizontal lines indicate the median. The p-value of the statistically significant difference between the two groups was calculated by Welch’s t-test; n = 4. g) Long-range PCR was performed on mtDNA isolated from the TA muscle of adult and aged wt or SAMHD1 −/− (ko) mice. FL, full-length product; the vertical line indicates the size range of mtDNA molecules with deletions. h) Kaplan–Meier survival curve for wt and SAMHD1 −/− (ko) mice. Comparison of the curves by the log-rank (Mantel–Cox) test confirmed no statistically significant difference between the genotypes. The sizes of the bands in the DNA ladder are indicated in kb. See also Fig. S5.
    Expand Long Template Pcr System, supplied by Roche, used in various techniques. Bioz Stars score: 92/100, based on 1024 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/expand long template pcr system/product/Roche
    Average 92 stars, based on 1024 article reviews
    Price from $9.99 to $1999.99
    expand long template pcr system - by Bioz Stars, 2021-01
    92/100 stars
      Buy from Supplier

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    Confirmation and characterization of the rearrangements . (A) Confirmation of the deletion of exons 1A/1B-2 by long-range PCR and sequencing of the breakpoints. (B) Confirmation of the deletion of exons 5–14 by long-range PCR and sequencing of the breakpoints. (C) Confirmation of the deletion of exons 11–12 by long-range PCR and sequencing of the breakpoints. Lanes 1+, 2+, carriers of the deletion; lane C-, negative control (wt); lane B, blank; lane M, marker (Ready-Load™ 1 Kb DNA Ladder, Invitrogen).

    Journal: BMC Medical Genetics

    Article Title: High occurrence of BRCA1 intragenic rearrangements in hereditary breast and ovarian cancer syndrome in the Czech Republic

    doi: 10.1186/1471-2350-8-32

    Figure Lengend Snippet: Confirmation and characterization of the rearrangements . (A) Confirmation of the deletion of exons 1A/1B-2 by long-range PCR and sequencing of the breakpoints. (B) Confirmation of the deletion of exons 5–14 by long-range PCR and sequencing of the breakpoints. (C) Confirmation of the deletion of exons 11–12 by long-range PCR and sequencing of the breakpoints. Lanes 1+, 2+, carriers of the deletion; lane C-, negative control (wt); lane B, blank; lane M, marker (Ready-Load™ 1 Kb DNA Ladder, Invitrogen).

    Article Snippet: Confirmation and characterization of the rearrangements Positive results detected by MLPA of two independently drawn samples of genomic DNA were confirmed by long-range PCR (Expand Long Template PCR System, Roche Applied Science), conducted in accordance with the manufacturer's instructions.

    Techniques: Polymerase Chain Reaction, Sequencing, Negative Control, Marker

    Confirmation and characterization of the rearrangements . (A) Confirmation of the deletion of exons 18–19 by long-range PCR and sequencing of the breakpoints. (B) Confirmation of the deletion of exon 20 and sequencing of the breakpoints.Lanes 1+, 2+, carriers of the deletion; lane C-, negative control (wt); lane B, blank; lane M, marker (Ready-Load™ 1 Kb DNA Ladder, Invitrogen).

    Journal: BMC Medical Genetics

    Article Title: High occurrence of BRCA1 intragenic rearrangements in hereditary breast and ovarian cancer syndrome in the Czech Republic

    doi: 10.1186/1471-2350-8-32

    Figure Lengend Snippet: Confirmation and characterization of the rearrangements . (A) Confirmation of the deletion of exons 18–19 by long-range PCR and sequencing of the breakpoints. (B) Confirmation of the deletion of exon 20 and sequencing of the breakpoints.Lanes 1+, 2+, carriers of the deletion; lane C-, negative control (wt); lane B, blank; lane M, marker (Ready-Load™ 1 Kb DNA Ladder, Invitrogen).

    Article Snippet: Confirmation and characterization of the rearrangements Positive results detected by MLPA of two independently drawn samples of genomic DNA were confirmed by long-range PCR (Expand Long Template PCR System, Roche Applied Science), conducted in accordance with the manufacturer's instructions.

    Techniques: Polymerase Chain Reaction, Sequencing, Negative Control, Marker

    Confirmation and characterization of the rearrangements . Confirmation of the deletion of the exons 21–22 by long-range PCR and sequencing of the breakpoints. The deletion/insertion event was characterized as g.77128_80906del3779ins236. Lanes 1+, 2+, carriers of the deletion; lane C-, negative control (wt); lane B, blank; lane M, marker (Ready-Load™ 1 Kb DNA Ladder, Invitrogen).

    Journal: BMC Medical Genetics

    Article Title: High occurrence of BRCA1 intragenic rearrangements in hereditary breast and ovarian cancer syndrome in the Czech Republic

    doi: 10.1186/1471-2350-8-32

    Figure Lengend Snippet: Confirmation and characterization of the rearrangements . Confirmation of the deletion of the exons 21–22 by long-range PCR and sequencing of the breakpoints. The deletion/insertion event was characterized as g.77128_80906del3779ins236. Lanes 1+, 2+, carriers of the deletion; lane C-, negative control (wt); lane B, blank; lane M, marker (Ready-Load™ 1 Kb DNA Ladder, Invitrogen).

    Article Snippet: Confirmation and characterization of the rearrangements Positive results detected by MLPA of two independently drawn samples of genomic DNA were confirmed by long-range PCR (Expand Long Template PCR System, Roche Applied Science), conducted in accordance with the manufacturer's instructions.

    Techniques: Polymerase Chain Reaction, Sequencing, Negative Control, Marker

    Deletion of SAMHD1 does not affect mtDNA stability. a) MtDNA copy number in the TA muscle of 5 or 6 wt (filled dots) and SAMHD1 −/− (open dots) 13-week-old (adult), 1-year-old (old adult), and 2-year-old (aged) animals was determined by qPCR and normalized to the value for adult wt mice. The mean for each group is indicated by a horizontal line. The p-values were calculated using Welch’s t-test; ns, non-significant. b) DNA isolated from embryos and from the TA muscle of pups, adults, 1-year-old (old) adults, or aged animals was linearized with SacI endonuclease and separated on a neutral gel. MtDNA was visualized as above. Full-length mtDNA is indicated (FL); the asterisk denotes a higher-migrating species resistant to cleavage. c) Long-range PCR to detect deletions in mtDNA from the TA muscle of wt mice of various ages. Full-length product is indicated (FL). Only minor species containing deletions are observed in the mtDNA from old adults and aged animals, as indicated by the vertical line on the right-hand side of the gel. d) Untreated or alkali-treated DNA from skeletal muscle of aged wt and SAMHD1 −/− (ko) mice was analyzed on a denaturing gel, and mtDNA was visualized using a COX1 probe. Each sample lane corresponds to an individual mouse, and dotted lines represent the median. e) The median length of the untreated mtDNA in samples from Fig. 5d is indicated by a horizontal line. The two groups were compared using Welch’s t-test (ns, non-significant; n = 4). f) The length difference between untreated and alkali-treated mtDNAs shown in Fig. 5d was used to compute the number of rNMPs per single strand of mtDNA. The horizontal lines indicate the median. The p-value of the statistically significant difference between the two groups was calculated by Welch’s t-test; n = 4. g) Long-range PCR was performed on mtDNA isolated from the TA muscle of adult and aged wt or SAMHD1 −/− (ko) mice. FL, full-length product; the vertical line indicates the size range of mtDNA molecules with deletions. h) Kaplan–Meier survival curve for wt and SAMHD1 −/− (ko) mice. Comparison of the curves by the log-rank (Mantel–Cox) test confirmed no statistically significant difference between the genotypes. The sizes of the bands in the DNA ladder are indicated in kb. See also Fig. S5.

    Journal: bioRxiv

    Article Title: The physiological level of rNMPs present in mtDNA does not compromise its stability

    doi: 10.1101/746719

    Figure Lengend Snippet: Deletion of SAMHD1 does not affect mtDNA stability. a) MtDNA copy number in the TA muscle of 5 or 6 wt (filled dots) and SAMHD1 −/− (open dots) 13-week-old (adult), 1-year-old (old adult), and 2-year-old (aged) animals was determined by qPCR and normalized to the value for adult wt mice. The mean for each group is indicated by a horizontal line. The p-values were calculated using Welch’s t-test; ns, non-significant. b) DNA isolated from embryos and from the TA muscle of pups, adults, 1-year-old (old) adults, or aged animals was linearized with SacI endonuclease and separated on a neutral gel. MtDNA was visualized as above. Full-length mtDNA is indicated (FL); the asterisk denotes a higher-migrating species resistant to cleavage. c) Long-range PCR to detect deletions in mtDNA from the TA muscle of wt mice of various ages. Full-length product is indicated (FL). Only minor species containing deletions are observed in the mtDNA from old adults and aged animals, as indicated by the vertical line on the right-hand side of the gel. d) Untreated or alkali-treated DNA from skeletal muscle of aged wt and SAMHD1 −/− (ko) mice was analyzed on a denaturing gel, and mtDNA was visualized using a COX1 probe. Each sample lane corresponds to an individual mouse, and dotted lines represent the median. e) The median length of the untreated mtDNA in samples from Fig. 5d is indicated by a horizontal line. The two groups were compared using Welch’s t-test (ns, non-significant; n = 4). f) The length difference between untreated and alkali-treated mtDNAs shown in Fig. 5d was used to compute the number of rNMPs per single strand of mtDNA. The horizontal lines indicate the median. The p-value of the statistically significant difference between the two groups was calculated by Welch’s t-test; n = 4. g) Long-range PCR was performed on mtDNA isolated from the TA muscle of adult and aged wt or SAMHD1 −/− (ko) mice. FL, full-length product; the vertical line indicates the size range of mtDNA molecules with deletions. h) Kaplan–Meier survival curve for wt and SAMHD1 −/− (ko) mice. Comparison of the curves by the log-rank (Mantel–Cox) test confirmed no statistically significant difference between the genotypes. The sizes of the bands in the DNA ladder are indicated in kb. See also Fig. S5.

    Article Snippet: MtDNA deletion analysis by long-range PCR The Expand Long Template PCR system (Roche) with forward and reverse primers at nt 2,478–2,512 and nt 1,933–1,906, respectively, was used to amplify a ~15,800 bp fragment of mouse mtDNA from 25 ng of total DNA.

    Techniques: Real-time Polymerase Chain Reaction, Mouse Assay, Isolation, Polymerase Chain Reaction

    Analysis of the PA2231-2245 gene cluster by RT-PCR of intergenic regions. Lane 1, size markers; lane 2, PCR amplification product of PA2230-2231 cluster with genomic DNA as the template (this serves as a control for RT-PCR of this region); lane 3, RT-PCR of PA2230 and -2231 (the lack of product indicates that PA2230 and -2231 are not cotranscribed); lane 4, size marker; lane 5, RT-PCR of PA2231 and -2232; lane 6, PA2232 and -2233; lane 7, PA2233 and -2234; lane 8, PA2234 and -2235; lane 9, PA2235 and -2236; lane 10, PA2236 and -2237; lane 11, PA2237 and -2238; lane 12, PA2238 and -2239; lane 13, PA2239 and -2240; lane 14, PA2240 and -2241; lane 15, PA2241 and -2242; lane 16, PA2242 and -2243; lane 17, PA2243 and -2244; lane 18, PA2244 and -2245. Control experiments without reverse transcriptase reactions prior to PCR were negative.

    Journal: Journal of Bacteriology

    Article Title: Putative Exopolysaccharide Synthesis Genes Influence Pseudomonas aeruginosa Biofilm Development

    doi: 10.1128/JB.186.14.4449-4456.2004

    Figure Lengend Snippet: Analysis of the PA2231-2245 gene cluster by RT-PCR of intergenic regions. Lane 1, size markers; lane 2, PCR amplification product of PA2230-2231 cluster with genomic DNA as the template (this serves as a control for RT-PCR of this region); lane 3, RT-PCR of PA2230 and -2231 (the lack of product indicates that PA2230 and -2231 are not cotranscribed); lane 4, size marker; lane 5, RT-PCR of PA2231 and -2232; lane 6, PA2232 and -2233; lane 7, PA2233 and -2234; lane 8, PA2234 and -2235; lane 9, PA2235 and -2236; lane 10, PA2236 and -2237; lane 11, PA2237 and -2238; lane 12, PA2238 and -2239; lane 13, PA2239 and -2240; lane 14, PA2240 and -2241; lane 15, PA2241 and -2242; lane 16, PA2242 and -2243; lane 17, PA2243 and -2244; lane 18, PA2244 and -2245. Control experiments without reverse transcriptase reactions prior to PCR were negative.

    Article Snippet: DNA contamination of the purified RNA was assessed by Expand long-template PCR using rplU primers.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Amplification, Marker

    Genomic distribution of L1 insertions. Outer rings show the density of detected insertion sites for reference (gray) and nonreference (black) L1s. The approximate locations of the 72 PCR-validated somatic insertions are indicated by dots inside the circle.

    Journal: Genome Research

    Article Title: Extensive somatic L1 retrotransposition in colorectal tumors

    doi: 10.1101/gr.145235.112

    Figure Lengend Snippet: Genomic distribution of L1 insertions. Outer rings show the density of detected insertion sites for reference (gray) and nonreference (black) L1s. The approximate locations of the 72 PCR-validated somatic insertions are indicated by dots inside the circle.

    Article Snippet: Long-range PCR to recover longer L1 insertions was performed with the Expand Long Template PCR System (Roche) according to the manufacturer's instructions in buffer 1, with 1 μL of 20 μM FS and ES primers each, and 25 ng of tumor DNA.

    Techniques: Polymerase Chain Reaction

    Analysis of factors influencing L1 activity. ( A ) L1 CpG promoter methylation status performed by quantitative bisulfite PCR analysis. (N) Normal tissue; (T) tumor tissue; (*) MSI. Replicates of four were done for each data point. (Error bars) Standard

    Journal: Genome Research

    Article Title: Extensive somatic L1 retrotransposition in colorectal tumors

    doi: 10.1101/gr.145235.112

    Figure Lengend Snippet: Analysis of factors influencing L1 activity. ( A ) L1 CpG promoter methylation status performed by quantitative bisulfite PCR analysis. (N) Normal tissue; (T) tumor tissue; (*) MSI. Replicates of four were done for each data point. (Error bars) Standard

    Article Snippet: Long-range PCR to recover longer L1 insertions was performed with the Expand Long Template PCR System (Roche) according to the manufacturer's instructions in buffer 1, with 1 μL of 20 μM FS and ES primers each, and 25 ng of tumor DNA.

    Techniques: Activity Assay, Methylation, Polymerase Chain Reaction

    PCR validation scheme of L1-seq results. ( A ) The three-step PCR validation scheme and location of primers used. Triangles symbolize TSD. ( B ) PCR validation of the 3′ junction (ins. 7). This insertion is in tumor 1 of the eight DNA samples that

    Journal: Genome Research

    Article Title: Extensive somatic L1 retrotransposition in colorectal tumors

    doi: 10.1101/gr.145235.112

    Figure Lengend Snippet: PCR validation scheme of L1-seq results. ( A ) The three-step PCR validation scheme and location of primers used. Triangles symbolize TSD. ( B ) PCR validation of the 3′ junction (ins. 7). This insertion is in tumor 1 of the eight DNA samples that

    Article Snippet: Long-range PCR to recover longer L1 insertions was performed with the Expand Long Template PCR System (Roche) according to the manufacturer's instructions in buffer 1, with 1 μL of 20 μM FS and ES primers each, and 25 ng of tumor DNA.

    Techniques: Polymerase Chain Reaction