pcr fragments  (Roche)


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    Structured Review

    Roche pcr fragments
    Characteristics of the circLMTK2 in GC cells. ( a ) The genomic loci of the LMTK2 gene and circLMTK2. The expression of circLMTK2 was detected by <t>qRT-PCR</t> and was validated by Sanger sequencing. The arrows represent divergent primers that bind to the genomic region of circLMTK2. ( b ) qRT-PCR analysis of circLMTK2 and LMTK2 mRNA after treatment with RNase R. ( c ) qRT-PCR analysis of circLMTK2 and LMTK2 mRNA after treatment with actinomycin D at the indicated time points. ( d ) qRT-PCR analysis of circLMTK2 and LMTK2 mRNA in either the cytoplasm or the nucleus. ( e ) <t>RNA</t> fluorescence in situ hybridization (FISH) for circLMTK2. The nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI). Scale bar, 5 μm
    Pcr Fragments, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr fragments/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcr fragments - by Bioz Stars, 2021-05
    86/100 stars

    Images

    1) Product Images from "circLMTK2 acts as a sponge of miR-150-5p and promotes proliferation and metastasis in gastric cancer"

    Article Title: circLMTK2 acts as a sponge of miR-150-5p and promotes proliferation and metastasis in gastric cancer

    Journal: Molecular Cancer

    doi: 10.1186/s12943-019-1081-4

    Characteristics of the circLMTK2 in GC cells. ( a ) The genomic loci of the LMTK2 gene and circLMTK2. The expression of circLMTK2 was detected by qRT-PCR and was validated by Sanger sequencing. The arrows represent divergent primers that bind to the genomic region of circLMTK2. ( b ) qRT-PCR analysis of circLMTK2 and LMTK2 mRNA after treatment with RNase R. ( c ) qRT-PCR analysis of circLMTK2 and LMTK2 mRNA after treatment with actinomycin D at the indicated time points. ( d ) qRT-PCR analysis of circLMTK2 and LMTK2 mRNA in either the cytoplasm or the nucleus. ( e ) RNA fluorescence in situ hybridization (FISH) for circLMTK2. The nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI). Scale bar, 5 μm
    Figure Legend Snippet: Characteristics of the circLMTK2 in GC cells. ( a ) The genomic loci of the LMTK2 gene and circLMTK2. The expression of circLMTK2 was detected by qRT-PCR and was validated by Sanger sequencing. The arrows represent divergent primers that bind to the genomic region of circLMTK2. ( b ) qRT-PCR analysis of circLMTK2 and LMTK2 mRNA after treatment with RNase R. ( c ) qRT-PCR analysis of circLMTK2 and LMTK2 mRNA after treatment with actinomycin D at the indicated time points. ( d ) qRT-PCR analysis of circLMTK2 and LMTK2 mRNA in either the cytoplasm or the nucleus. ( e ) RNA fluorescence in situ hybridization (FISH) for circLMTK2. The nuclei were stained with 4,6-diamidino-2-phenylindole (DAPI). Scale bar, 5 μm

    Techniques Used: Expressing, Quantitative RT-PCR, Sequencing, Fluorescence, In Situ Hybridization, Fluorescence In Situ Hybridization, Staining

    2) Product Images from "A PCR-Based Method for RNA Probes and Applications in Neuroscience"

    Article Title: A PCR-Based Method for RNA Probes and Applications in Neuroscience

    Journal: Frontiers in Neuroscience

    doi: 10.3389/fnins.2018.00266

    Preparation and visualization of SST probe in mouse brain sections. Two-step polymerase chain reaction (PCR) amplifications were performed and PCR products were examined by agarose gel electrophoresis. (A) Products of SST from the first PCR. (B) Products of SST containing the T7 promoter from the second PCR. (C) In vitro -transcribed RNA probe of SST. (D) Staining for SST mRNA expression revealed the effect of tissue permeability on signal intensity. The left images show treatment with 1 × PBST (1% Tween-20 in 0.01 M PBS) for 20 min at room temperature (RT); the middle images show treatment with 2 μg/ml proteinase K at RT; and the right images show treatment with 2 μg/ml proteinase K at 37°C. The upper images were captured using 10 × magnification (scale bar = 500 μm), and the bottom images were captured using 20 × magnification (scale bar = 100 μm). (E) Sections were stained with different concentrations of the SST probe at 0, 0.5, 2, and 4 μg/ml, respectively. Images were captured using 10 × magnification (scale bar = 200 μm). SST, somatostatin.
    Figure Legend Snippet: Preparation and visualization of SST probe in mouse brain sections. Two-step polymerase chain reaction (PCR) amplifications were performed and PCR products were examined by agarose gel electrophoresis. (A) Products of SST from the first PCR. (B) Products of SST containing the T7 promoter from the second PCR. (C) In vitro -transcribed RNA probe of SST. (D) Staining for SST mRNA expression revealed the effect of tissue permeability on signal intensity. The left images show treatment with 1 × PBST (1% Tween-20 in 0.01 M PBS) for 20 min at room temperature (RT); the middle images show treatment with 2 μg/ml proteinase K at RT; and the right images show treatment with 2 μg/ml proteinase K at 37°C. The upper images were captured using 10 × magnification (scale bar = 500 μm), and the bottom images were captured using 20 × magnification (scale bar = 100 μm). (E) Sections were stained with different concentrations of the SST probe at 0, 0.5, 2, and 4 μg/ml, respectively. Images were captured using 10 × magnification (scale bar = 200 μm). SST, somatostatin.

    Techniques Used: Polymerase Chain Reaction, Agarose Gel Electrophoresis, In Vitro, Staining, Expressing, Permeability

    3) Product Images from "XX/XY System of Sex Determination in the Geophilomorph Centipede Strigamia maritima"

    Article Title: XX/XY System of Sex Determination in the Geophilomorph Centipede Strigamia maritima

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0150292

    Identification of the X chromosome in the Strigamia karyotype by FISH with a set of X-chromosome derived DNA probes. (A) The relative positions and sizes of the five PCR fragments, distributed on two different X-linked scaffolds, which were used to produce the X-probes. Two of the fragments are located on scf7180001248200 and three on scf7180001248049. The genomic distance between these scaffolds is not known. The black line depicts the scaffold. The numbers above the line indicate the number of bases, starting at 1 on the left hand side. Green boxes represent the length and relative position of the PCR products, numbered arbitrarily from 1 to 5. (B) A mitotic metaphase chromosome spread prepared from a single embryo. Hybridization signals of the X-probes identify a middle-sized element in the Strigamia karyotype as the X chromosome. As there are two chromosomes with the X-probe signals, we infer that this chromosome spread is derived from a female embryo (XX). (C) Two Strigamia karyotypes constructed from the mitotic metaphases of embryonic cells. They are derived from different embryos. Upper panel: karyotype derived from the female metaphase shown in (B). Lower panel: karyotype derived from an inverted image of a DAPI-stained metaphase of unknown sex. It is the same as that shown in Fig 1A . We infer that the pair of sex chromosomes represents the 4 th pair of chromosomes by size (asterisks). (D, E, F) Meiotic chromosome spreads, prepared from sub-adult male testes. (D) Late zygotene complement showing a clump of incompletely paired bivalents. The X-probes label the longer chromosome of a partially paired bivalent, as schematically illustrated in (D’). We thus infer that this is the X chromosome, and that the other shorter chromosome, without hybridization signals, is the Y chromosome. The X and Y chromosomes are only paired at the distal part of the X chromosome, with a large proximal part unpaired. (E) A particularly clear and well-spread XY bivalent at a similar stage to (D). It shows hybridization signals of X-probes on the unpaired proximal part of the X chromosome, while the Y chromosome is completely paired except for the DAPI-highlighted centromere (see schematic drawing below the XY bivalent). (F) Pachytene complement showing 8 bivalents, each with DAPI-highlighted centromeric chromatin. X-probe hybridization signals are visible on the unpaired segment of the longer chromosome, near the centromere (see schematic drawing on the right-hand side). The X and Y chromosomes now appear almost equal in length in the bivalent. Scale bar is equal to 5 μm in (B) and 10 μm in (D, E, F). Chromosomes were counterstained with DAPI (blue). Arrowheads indicate hybridization signals of the digoxigenin-labelled X-probes (green); arrows indicate a pair of the largest chromosomes (B) or the largest bivalent (F).
    Figure Legend Snippet: Identification of the X chromosome in the Strigamia karyotype by FISH with a set of X-chromosome derived DNA probes. (A) The relative positions and sizes of the five PCR fragments, distributed on two different X-linked scaffolds, which were used to produce the X-probes. Two of the fragments are located on scf7180001248200 and three on scf7180001248049. The genomic distance between these scaffolds is not known. The black line depicts the scaffold. The numbers above the line indicate the number of bases, starting at 1 on the left hand side. Green boxes represent the length and relative position of the PCR products, numbered arbitrarily from 1 to 5. (B) A mitotic metaphase chromosome spread prepared from a single embryo. Hybridization signals of the X-probes identify a middle-sized element in the Strigamia karyotype as the X chromosome. As there are two chromosomes with the X-probe signals, we infer that this chromosome spread is derived from a female embryo (XX). (C) Two Strigamia karyotypes constructed from the mitotic metaphases of embryonic cells. They are derived from different embryos. Upper panel: karyotype derived from the female metaphase shown in (B). Lower panel: karyotype derived from an inverted image of a DAPI-stained metaphase of unknown sex. It is the same as that shown in Fig 1A . We infer that the pair of sex chromosomes represents the 4 th pair of chromosomes by size (asterisks). (D, E, F) Meiotic chromosome spreads, prepared from sub-adult male testes. (D) Late zygotene complement showing a clump of incompletely paired bivalents. The X-probes label the longer chromosome of a partially paired bivalent, as schematically illustrated in (D’). We thus infer that this is the X chromosome, and that the other shorter chromosome, without hybridization signals, is the Y chromosome. The X and Y chromosomes are only paired at the distal part of the X chromosome, with a large proximal part unpaired. (E) A particularly clear and well-spread XY bivalent at a similar stage to (D). It shows hybridization signals of X-probes on the unpaired proximal part of the X chromosome, while the Y chromosome is completely paired except for the DAPI-highlighted centromere (see schematic drawing below the XY bivalent). (F) Pachytene complement showing 8 bivalents, each with DAPI-highlighted centromeric chromatin. X-probe hybridization signals are visible on the unpaired segment of the longer chromosome, near the centromere (see schematic drawing on the right-hand side). The X and Y chromosomes now appear almost equal in length in the bivalent. Scale bar is equal to 5 μm in (B) and 10 μm in (D, E, F). Chromosomes were counterstained with DAPI (blue). Arrowheads indicate hybridization signals of the digoxigenin-labelled X-probes (green); arrows indicate a pair of the largest chromosomes (B) or the largest bivalent (F).

    Techniques Used: Fluorescence In Situ Hybridization, Derivative Assay, Polymerase Chain Reaction, Hybridization, Construct, Staining

    Related Articles

    Polymerase Chain Reaction:

    Article Title: Depletion of SNAP-23 and Syntaxin 4 alters lipid droplet homeostasis during Chlamydia infection
    Article Snippet: Note that both rat Syntaxin 3 and Syntaxin 4 are 97.22% and 89.23% identical to human Syntaxin 3 and Syntaxin 4, respectively (Fig. S10) and are commonly used to replace human Syntaxins [ , ]. .. PCR fragments were cloned into the EcoRI and XhoI sites of pCMV-Tag2B (gift from Paul Roche, NIH), which was modified to contain a 3xFLAG tag instead of a single FLAG tag. .. DNA sequences were confirmed by Sanger sequencing (GenScript).

    Article Title: Analyses of the Evolutionary Distribution of Salmonella Translocated Effectors
    Article Snippet: Gene-specific hybridization probes for STE genes were obtained by PCR using the primers indicated in Table and the legend to Fig. with the genomic DNA of S. enterica subspecies I serotype Typhimurium (serovar Typhimurium) strain ATCC 14028 as the template. .. PCR fragments were labeled by using the DIG DNA Labeling Kit (Roche). .. Hybridization was carried out at 42°C overnight with 20% (vol/vol) formamide in 1× hybridization buffer (250 mM NaHPO4 , 1 mM EDTA, 7% sodium dodecyl sulfate).

    Article Title: An Atypical Proprotein Convertase in Giardia lamblia Differentiation
    Article Snippet: The vectors were de-phosphorylated using shrimp alkaline phosphatase (Promega; Madison, WI) by standard methods and purified using QIAquick (Qiagen). .. The digested vectors and PCR fragments were ligated (T4 DNA ligase; Roche, Nutley, NJ) overnight at 4 °C, transformed into JM109 competent cells (Promega), and plasmids screened by size for automated sequencing of clones containing full-length gSPC. .. WB clone C6 trophozoites were electroporated with 50 μg of plasmid containing gSPC-AU1 or gSPC-HA and were selected and maintained by puromycin resistance [ ].

    Article Title: Decoding Hematopoietic Specificity in the Helix-Loop-Helix Domain of the Transcription Factor SCL/Tal-1
    Article Snippet: .. PCR fragments containing a T7 promoter sequence at their 5′ end were generated from Flag-tagged SCL (wt and mutant) constructs and E12 cDNA by using the Expand long-template polymerase (Roche). .. Equal amounts of SCL (wt or mutant) and E12 PCR products were mixed and used as templates for coupled in vitro transcription-translation with the TNT T7 Quick for PCR DNA (Promega).

    Article Title: Morphological and stage-specific transcriptome analyses reveal distinct regulatory programs underlying yam (Dioscorea alata L.) bulbil growth
    Article Snippet: Gene-specific fragments for probe synthesis were amplified by PCR using designed primers: 5'-GAAGAGCACCATGCTGTGAG-3' and 5'-TAATACGACTCACTATAGGGCCACATCTCAGCAATCCAG-3' for MYB (Gene ID: c126446.graph-c0), 5'-TGGAGAGCCTTTGATCGGTT-3' and 5'-TAATACGACTCACTATAGGGCCACTGCTCTAAACGAAGG-3' for WRKY (c125026.graph_c1), and 5'-AGTGCATTACCTCTGCCGGA-3' and 5'-TAATACGACTCACTATAGGTACCTGGCAATTCCCAAGGA-3' for NAC (c116834.graph_c0). .. The resulting PCR fragments were used as templates for synthesis of digoxigenin (DIG)-labeled antisense and sense riboprobes with the T7/SP6 riboprobe and a DIG-RNA labeling mix (Roche). ..

    Article Title: Genotyping of Hepatitis C Virus Isolates using CLIP Sequencing
    Article Snippet: .. This technique utilizes PCR fragments previously generated by the diagnostic Roche AMPLICOR HCV test, which are subsequently subjected to simultaneous PCR amplification and direct sequencing (CLIP sequencing) of the 5′ noncoding region (5′NCR). .. HCV isolates from 100 randomly chosen patients were genotyped by both the TRUGENE HCV 5′NC genotyping kit and DNA enzyme immunoassay (DEIA).

    Article Title: Nuclear Accumulation of ?-Catenin Protein in Chemically Induced Rat Nephroblastomas
    Article Snippet: .. PCR fragments were purified using High Pure PCR Purification Kit (Roche Diagnostics, Basel, Switzerland), essentially as recommended by the manufacturer. .. Sequencing reactions were set up using 30 ng of purified PCR product and 10 pmol of sequencing primer in a total reaction volume of 10 μl following a dye terminator protocol (Big Dye, Applied Biosystems, Darmstadt, Germany).

    Clone Assay:

    Article Title: Depletion of SNAP-23 and Syntaxin 4 alters lipid droplet homeostasis during Chlamydia infection
    Article Snippet: Note that both rat Syntaxin 3 and Syntaxin 4 are 97.22% and 89.23% identical to human Syntaxin 3 and Syntaxin 4, respectively (Fig. S10) and are commonly used to replace human Syntaxins [ , ]. .. PCR fragments were cloned into the EcoRI and XhoI sites of pCMV-Tag2B (gift from Paul Roche, NIH), which was modified to contain a 3xFLAG tag instead of a single FLAG tag. .. DNA sequences were confirmed by Sanger sequencing (GenScript).

    Article Title: An Atypical Proprotein Convertase in Giardia lamblia Differentiation
    Article Snippet: The vectors were de-phosphorylated using shrimp alkaline phosphatase (Promega; Madison, WI) by standard methods and purified using QIAquick (Qiagen). .. The digested vectors and PCR fragments were ligated (T4 DNA ligase; Roche, Nutley, NJ) overnight at 4 °C, transformed into JM109 competent cells (Promega), and plasmids screened by size for automated sequencing of clones containing full-length gSPC. .. WB clone C6 trophozoites were electroporated with 50 μg of plasmid containing gSPC-AU1 or gSPC-HA and were selected and maintained by puromycin resistance [ ].

    Modification:

    Article Title: Depletion of SNAP-23 and Syntaxin 4 alters lipid droplet homeostasis during Chlamydia infection
    Article Snippet: Note that both rat Syntaxin 3 and Syntaxin 4 are 97.22% and 89.23% identical to human Syntaxin 3 and Syntaxin 4, respectively (Fig. S10) and are commonly used to replace human Syntaxins [ , ]. .. PCR fragments were cloned into the EcoRI and XhoI sites of pCMV-Tag2B (gift from Paul Roche, NIH), which was modified to contain a 3xFLAG tag instead of a single FLAG tag. .. DNA sequences were confirmed by Sanger sequencing (GenScript).

    FLAG-tag:

    Article Title: Depletion of SNAP-23 and Syntaxin 4 alters lipid droplet homeostasis during Chlamydia infection
    Article Snippet: Note that both rat Syntaxin 3 and Syntaxin 4 are 97.22% and 89.23% identical to human Syntaxin 3 and Syntaxin 4, respectively (Fig. S10) and are commonly used to replace human Syntaxins [ , ]. .. PCR fragments were cloned into the EcoRI and XhoI sites of pCMV-Tag2B (gift from Paul Roche, NIH), which was modified to contain a 3xFLAG tag instead of a single FLAG tag. .. DNA sequences were confirmed by Sanger sequencing (GenScript).

    Labeling:

    Article Title: Analyses of the Evolutionary Distribution of Salmonella Translocated Effectors
    Article Snippet: Gene-specific hybridization probes for STE genes were obtained by PCR using the primers indicated in Table and the legend to Fig. with the genomic DNA of S. enterica subspecies I serotype Typhimurium (serovar Typhimurium) strain ATCC 14028 as the template. .. PCR fragments were labeled by using the DIG DNA Labeling Kit (Roche). .. Hybridization was carried out at 42°C overnight with 20% (vol/vol) formamide in 1× hybridization buffer (250 mM NaHPO4 , 1 mM EDTA, 7% sodium dodecyl sulfate).

    Article Title: Morphological and stage-specific transcriptome analyses reveal distinct regulatory programs underlying yam (Dioscorea alata L.) bulbil growth
    Article Snippet: Gene-specific fragments for probe synthesis were amplified by PCR using designed primers: 5'-GAAGAGCACCATGCTGTGAG-3' and 5'-TAATACGACTCACTATAGGGCCACATCTCAGCAATCCAG-3' for MYB (Gene ID: c126446.graph-c0), 5'-TGGAGAGCCTTTGATCGGTT-3' and 5'-TAATACGACTCACTATAGGGCCACTGCTCTAAACGAAGG-3' for WRKY (c125026.graph_c1), and 5'-AGTGCATTACCTCTGCCGGA-3' and 5'-TAATACGACTCACTATAGGTACCTGGCAATTCCCAAGGA-3' for NAC (c116834.graph_c0). .. The resulting PCR fragments were used as templates for synthesis of digoxigenin (DIG)-labeled antisense and sense riboprobes with the T7/SP6 riboprobe and a DIG-RNA labeling mix (Roche). ..

    DNA Labeling:

    Article Title: Analyses of the Evolutionary Distribution of Salmonella Translocated Effectors
    Article Snippet: Gene-specific hybridization probes for STE genes were obtained by PCR using the primers indicated in Table and the legend to Fig. with the genomic DNA of S. enterica subspecies I serotype Typhimurium (serovar Typhimurium) strain ATCC 14028 as the template. .. PCR fragments were labeled by using the DIG DNA Labeling Kit (Roche). .. Hybridization was carried out at 42°C overnight with 20% (vol/vol) formamide in 1× hybridization buffer (250 mM NaHPO4 , 1 mM EDTA, 7% sodium dodecyl sulfate).

    Transformation Assay:

    Article Title: An Atypical Proprotein Convertase in Giardia lamblia Differentiation
    Article Snippet: The vectors were de-phosphorylated using shrimp alkaline phosphatase (Promega; Madison, WI) by standard methods and purified using QIAquick (Qiagen). .. The digested vectors and PCR fragments were ligated (T4 DNA ligase; Roche, Nutley, NJ) overnight at 4 °C, transformed into JM109 competent cells (Promega), and plasmids screened by size for automated sequencing of clones containing full-length gSPC. .. WB clone C6 trophozoites were electroporated with 50 μg of plasmid containing gSPC-AU1 or gSPC-HA and were selected and maintained by puromycin resistance [ ].

    Sequencing:

    Article Title: An Atypical Proprotein Convertase in Giardia lamblia Differentiation
    Article Snippet: The vectors were de-phosphorylated using shrimp alkaline phosphatase (Promega; Madison, WI) by standard methods and purified using QIAquick (Qiagen). .. The digested vectors and PCR fragments were ligated (T4 DNA ligase; Roche, Nutley, NJ) overnight at 4 °C, transformed into JM109 competent cells (Promega), and plasmids screened by size for automated sequencing of clones containing full-length gSPC. .. WB clone C6 trophozoites were electroporated with 50 μg of plasmid containing gSPC-AU1 or gSPC-HA and were selected and maintained by puromycin resistance [ ].

    Article Title: Decoding Hematopoietic Specificity in the Helix-Loop-Helix Domain of the Transcription Factor SCL/Tal-1
    Article Snippet: .. PCR fragments containing a T7 promoter sequence at their 5′ end were generated from Flag-tagged SCL (wt and mutant) constructs and E12 cDNA by using the Expand long-template polymerase (Roche). .. Equal amounts of SCL (wt or mutant) and E12 PCR products were mixed and used as templates for coupled in vitro transcription-translation with the TNT T7 Quick for PCR DNA (Promega).

    Article Title: Genotyping of Hepatitis C Virus Isolates using CLIP Sequencing
    Article Snippet: .. This technique utilizes PCR fragments previously generated by the diagnostic Roche AMPLICOR HCV test, which are subsequently subjected to simultaneous PCR amplification and direct sequencing (CLIP sequencing) of the 5′ noncoding region (5′NCR). .. HCV isolates from 100 randomly chosen patients were genotyped by both the TRUGENE HCV 5′NC genotyping kit and DNA enzyme immunoassay (DEIA).

    Generated:

    Article Title: Decoding Hematopoietic Specificity in the Helix-Loop-Helix Domain of the Transcription Factor SCL/Tal-1
    Article Snippet: .. PCR fragments containing a T7 promoter sequence at their 5′ end were generated from Flag-tagged SCL (wt and mutant) constructs and E12 cDNA by using the Expand long-template polymerase (Roche). .. Equal amounts of SCL (wt or mutant) and E12 PCR products were mixed and used as templates for coupled in vitro transcription-translation with the TNT T7 Quick for PCR DNA (Promega).

    Article Title: Genotyping of Hepatitis C Virus Isolates using CLIP Sequencing
    Article Snippet: .. This technique utilizes PCR fragments previously generated by the diagnostic Roche AMPLICOR HCV test, which are subsequently subjected to simultaneous PCR amplification and direct sequencing (CLIP sequencing) of the 5′ noncoding region (5′NCR). .. HCV isolates from 100 randomly chosen patients were genotyped by both the TRUGENE HCV 5′NC genotyping kit and DNA enzyme immunoassay (DEIA).

    Mutagenesis:

    Article Title: Decoding Hematopoietic Specificity in the Helix-Loop-Helix Domain of the Transcription Factor SCL/Tal-1
    Article Snippet: .. PCR fragments containing a T7 promoter sequence at their 5′ end were generated from Flag-tagged SCL (wt and mutant) constructs and E12 cDNA by using the Expand long-template polymerase (Roche). .. Equal amounts of SCL (wt or mutant) and E12 PCR products were mixed and used as templates for coupled in vitro transcription-translation with the TNT T7 Quick for PCR DNA (Promega).

    Construct:

    Article Title: Decoding Hematopoietic Specificity in the Helix-Loop-Helix Domain of the Transcription Factor SCL/Tal-1
    Article Snippet: .. PCR fragments containing a T7 promoter sequence at their 5′ end were generated from Flag-tagged SCL (wt and mutant) constructs and E12 cDNA by using the Expand long-template polymerase (Roche). .. Equal amounts of SCL (wt or mutant) and E12 PCR products were mixed and used as templates for coupled in vitro transcription-translation with the TNT T7 Quick for PCR DNA (Promega).

    Amplification:

    Article Title: A Mitogenomic Phylogeny of Living Primates
    Article Snippet: .. Two overlapping PCR fragments with sizes of 8 kb (primers 5’-GGCTTTCTCAACTTTTAAAGGATA-3’ ; 5’-TGTCCTGATCCAACATCGAG-3’ ) and 10 kb (primers 5’-CCGTGCAAAGGTAGCATAATC-3’ ; 5’-TTACTTTTATTTGGAGTTGCACCA-3’ ), respectively, that cover the entire mt genome were amplified using the Expand Long Range dNTPack (Roche). ..

    Article Title: Genotyping of Hepatitis C Virus Isolates using CLIP Sequencing
    Article Snippet: .. This technique utilizes PCR fragments previously generated by the diagnostic Roche AMPLICOR HCV test, which are subsequently subjected to simultaneous PCR amplification and direct sequencing (CLIP sequencing) of the 5′ noncoding region (5′NCR). .. HCV isolates from 100 randomly chosen patients were genotyped by both the TRUGENE HCV 5′NC genotyping kit and DNA enzyme immunoassay (DEIA).

    Diagnostic Assay:

    Article Title: Genotyping of Hepatitis C Virus Isolates using CLIP Sequencing
    Article Snippet: .. This technique utilizes PCR fragments previously generated by the diagnostic Roche AMPLICOR HCV test, which are subsequently subjected to simultaneous PCR amplification and direct sequencing (CLIP sequencing) of the 5′ noncoding region (5′NCR). .. HCV isolates from 100 randomly chosen patients were genotyped by both the TRUGENE HCV 5′NC genotyping kit and DNA enzyme immunoassay (DEIA).

    Cross-linking Immunoprecipitation:

    Article Title: Genotyping of Hepatitis C Virus Isolates using CLIP Sequencing
    Article Snippet: .. This technique utilizes PCR fragments previously generated by the diagnostic Roche AMPLICOR HCV test, which are subsequently subjected to simultaneous PCR amplification and direct sequencing (CLIP sequencing) of the 5′ noncoding region (5′NCR). .. HCV isolates from 100 randomly chosen patients were genotyped by both the TRUGENE HCV 5′NC genotyping kit and DNA enzyme immunoassay (DEIA).

    Purification:

    Article Title: Nuclear Accumulation of ?-Catenin Protein in Chemically Induced Rat Nephroblastomas
    Article Snippet: .. PCR fragments were purified using High Pure PCR Purification Kit (Roche Diagnostics, Basel, Switzerland), essentially as recommended by the manufacturer. .. Sequencing reactions were set up using 30 ng of purified PCR product and 10 pmol of sequencing primer in a total reaction volume of 10 μl following a dye terminator protocol (Big Dye, Applied Biosystems, Darmstadt, Germany).

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    Roche pcr fragment
    The transcription factors NFE2 and MTF-1 may be involved in the differential expression of <t>Abcc6</t> in the liver of β-thalassemia mouse model ( Hbb th3/+ ) mice. Soluble chromatin prepared from the liver of wild-type (WT) ( A , upper panels ) and Hbb th3/+ mice ( A , lower panels ) was immunoprecipitated with an antibody raised against the indicated transcription factor. Negative control immunoprecipitations were performed with a nonimmunized control serum (NoAb). Chromatin preparations were analyzed for specific enrichment by <t>PCR</t> ( A ) and measured by quantitative PCR ( B ) using a pair of primers covering the promoter region of interest. For a positive control, chromatin extracts before immunoprecipitation (Input) was amplified in parallel. Standard errors are shown. * P
    Pcr Fragment, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr fragment/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcr fragment - by Bioz Stars, 2021-05
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    Roche pcr fragments
    RacR binds to the ggt promoter region as shown by electrophoretic mobility shift assays (EMSA) . <t>DIG-labeled</t> <t>PCR</t> fragments (~50 fmol) containing the ggt, phoX , or Cj0200c promoter regions were incubated with RacR as indicated. (A) Influence of the phosphorylation of RacR on the binding to the ggt promoter. RacR was phosphorylated by RacScyto in the presence of ATP. (B) EMSA of the ggt, phoX , and Cj0200c promoter regions with phosphorylated RacR protein. The phoX and Cj0200c promoter regions were used as negative controls. RacR-P, phosphorylated RacR.
    Pcr Fragments, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr fragments/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcr fragments - by Bioz Stars, 2021-05
    86/100 stars
      Buy from Supplier

    Image Search Results


    The transcription factors NFE2 and MTF-1 may be involved in the differential expression of Abcc6 in the liver of β-thalassemia mouse model ( Hbb th3/+ ) mice. Soluble chromatin prepared from the liver of wild-type (WT) ( A , upper panels ) and Hbb th3/+ mice ( A , lower panels ) was immunoprecipitated with an antibody raised against the indicated transcription factor. Negative control immunoprecipitations were performed with a nonimmunized control serum (NoAb). Chromatin preparations were analyzed for specific enrichment by PCR ( A ) and measured by quantitative PCR ( B ) using a pair of primers covering the promoter region of interest. For a positive control, chromatin extracts before immunoprecipitation (Input) was amplified in parallel. Standard errors are shown. * P

    Journal: The American Journal of Pathology

    Article Title: A Mouse Model of ?-Thalassemia Shows a Liver-Specific Down-Regulation of Abcc6 Expression

    doi: 10.1016/j.ajpath.2010.10.004

    Figure Lengend Snippet: The transcription factors NFE2 and MTF-1 may be involved in the differential expression of Abcc6 in the liver of β-thalassemia mouse model ( Hbb th3/+ ) mice. Soluble chromatin prepared from the liver of wild-type (WT) ( A , upper panels ) and Hbb th3/+ mice ( A , lower panels ) was immunoprecipitated with an antibody raised against the indicated transcription factor. Negative control immunoprecipitations were performed with a nonimmunized control serum (NoAb). Chromatin preparations were analyzed for specific enrichment by PCR ( A ) and measured by quantitative PCR ( B ) using a pair of primers covering the promoter region of interest. For a positive control, chromatin extracts before immunoprecipitation (Input) was amplified in parallel. Standard errors are shown. * P

    Article Snippet: This PCR fragment corresponding to the mouse Abcc6 promoter region was digested with KpnI and subsequently labeled with biotin using Bio-16-dUTP (Roche) and a terminal transferase (New England Biolabs, Beverly, MA).

    Techniques: Expressing, Mouse Assay, Immunoprecipitation, Negative Control, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Positive Control, Amplification

    Mutation in the RP1 gene due to nucleotide substitution . A. Pedigree, restriction analysis, and direct sequencing of Spanish families showing the R677X mutation. The 2177 C→T substitution abolishes the TaqI restriction site. M is the DNA marker consisting of a 100 bp ladder and U/D is the undigested DNA PCR fragment. B . Pedigree, DGGE and direct sequencing of the family carrying the Q686X mutation in the RP1 gene. Affected individuals, asymptomatic carriers and non-carriers of the mutation are represented by solid symbols, symbols with an internal dot and open symbols, respectively.

    Journal: BMC Medical Genetics

    Article Title: Three novel and the common Arg677Ter RP1 protein truncating mutations causing autosomal dominant retinitis pigmentosa in a Spanish population

    doi: 10.1186/1471-2350-7-35

    Figure Lengend Snippet: Mutation in the RP1 gene due to nucleotide substitution . A. Pedigree, restriction analysis, and direct sequencing of Spanish families showing the R677X mutation. The 2177 C→T substitution abolishes the TaqI restriction site. M is the DNA marker consisting of a 100 bp ladder and U/D is the undigested DNA PCR fragment. B . Pedigree, DGGE and direct sequencing of the family carrying the Q686X mutation in the RP1 gene. Affected individuals, asymptomatic carriers and non-carriers of the mutation are represented by solid symbols, symbols with an internal dot and open symbols, respectively.

    Article Snippet: The DNA from the PCR fragment containing the R677X mutation was digested with the endonuclease TaqI (Roche, Barcelona, Spain), according to the manufacturer's specifications.

    Techniques: Mutagenesis, Sequencing, Marker, Polymerase Chain Reaction, Denaturing Gradient Gel Electrophoresis

    Schematic representation of the CYP2E1 gene showing the polymorphism location and the design of the recRNA. For the genomic structure, exons, indicated by boxes, are numbered, and introns are represented by horizontal lines. The arrow indicates the transcription start site, and the Rsa I/ Pst I polymorphism sites are denoted. To amplify the region of the CYP2E1 gene encompassing exons 2–5, we used forward primer T7CYP2E1 on the junction of exons 2–3 and reverse primer RecRNA in exon 5. The presence of the T7 promoter sequence at the 5′ end of the forward primer enabled the recRNA to be produced by in vitro transcription. Cellular RNA and recRNA were co-reverse transcribed and amplified simultaneously with the CYP2E1F and CYP2E1R primers. Because the 20 bp stretch incorporated at the 3′ end of the primer RecRNA (thin line on standard) is complementary to primer CYP2E1, the RT-PCR reaction yielded products of 453 bp from the cellular RNA and of 382 bp from the recRNA. Both PCR products had the same DNA sequence, except for the 71 bp deletion in the middle of the standard fragment.

    Journal: Environmental Health Perspectives

    Article Title: Occupational Toluene Exposure Induces Cytochrome P450 2E1 mRNA Expression in Peripheral Lymphocytes

    doi: 10.1289/ehp.8192

    Figure Lengend Snippet: Schematic representation of the CYP2E1 gene showing the polymorphism location and the design of the recRNA. For the genomic structure, exons, indicated by boxes, are numbered, and introns are represented by horizontal lines. The arrow indicates the transcription start site, and the Rsa I/ Pst I polymorphism sites are denoted. To amplify the region of the CYP2E1 gene encompassing exons 2–5, we used forward primer T7CYP2E1 on the junction of exons 2–3 and reverse primer RecRNA in exon 5. The presence of the T7 promoter sequence at the 5′ end of the forward primer enabled the recRNA to be produced by in vitro transcription. Cellular RNA and recRNA were co-reverse transcribed and amplified simultaneously with the CYP2E1F and CYP2E1R primers. Because the 20 bp stretch incorporated at the 3′ end of the primer RecRNA (thin line on standard) is complementary to primer CYP2E1, the RT-PCR reaction yielded products of 453 bp from the cellular RNA and of 382 bp from the recRNA. Both PCR products had the same DNA sequence, except for the 71 bp deletion in the middle of the standard fragment.

    Article Snippet: In the second step, recRNA derived from the PCR fragment containing the T7 promoter sequence was prepared by in vitro transcription with a T7 Transcription Kit (Roche Diagnostics GmbH, Mannheim, Germany), according to the manufacturer’s instructions.

    Techniques: Sequencing, Produced, In Vitro, Amplification, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction

    Reliability of the competitive RT-PCR assay for CYP2E1 transcript. ( A ) A fixed amount of human lymphocyte total RNA (5 μg) was incubated with decreasing amounts of recRNA (lane 1, 1 ng; lane 2, 100 pg; lane 3, 10 pg; lane 4, 1 pg; lane 5, 100 fg; lane 6, 10 fg; lane 7, 1 fg; lane 8, without recRNA. The expected PCR products of 453 and 382 bp correspond to endogenous CYP2E1 mRNA and recRNA, respectively. ( B ) A fixed amount of recRNA (1 pg) was incubated with decreasing amounts of human lymphocyte total RNA. Lane 1, 10 μg; lane 2, 7.5 μg; lane 3, 5 μg; lane 4, 2.5 μg; lane 5, 1 μg; lane 6, without RNA. ( C ) Representative gel of the competitive RT-PCR assay showing the CYP2E1 mRNA expression of different subjects (5 μg of total RNA and 10 pg of recRNA; lanes 1–5). Abbreviations: –, control with RNA sample omitted; M, DNA ladder.

    Journal: Environmental Health Perspectives

    Article Title: Occupational Toluene Exposure Induces Cytochrome P450 2E1 mRNA Expression in Peripheral Lymphocytes

    doi: 10.1289/ehp.8192

    Figure Lengend Snippet: Reliability of the competitive RT-PCR assay for CYP2E1 transcript. ( A ) A fixed amount of human lymphocyte total RNA (5 μg) was incubated with decreasing amounts of recRNA (lane 1, 1 ng; lane 2, 100 pg; lane 3, 10 pg; lane 4, 1 pg; lane 5, 100 fg; lane 6, 10 fg; lane 7, 1 fg; lane 8, without recRNA. The expected PCR products of 453 and 382 bp correspond to endogenous CYP2E1 mRNA and recRNA, respectively. ( B ) A fixed amount of recRNA (1 pg) was incubated with decreasing amounts of human lymphocyte total RNA. Lane 1, 10 μg; lane 2, 7.5 μg; lane 3, 5 μg; lane 4, 2.5 μg; lane 5, 1 μg; lane 6, without RNA. ( C ) Representative gel of the competitive RT-PCR assay showing the CYP2E1 mRNA expression of different subjects (5 μg of total RNA and 10 pg of recRNA; lanes 1–5). Abbreviations: –, control with RNA sample omitted; M, DNA ladder.

    Article Snippet: In the second step, recRNA derived from the PCR fragment containing the T7 promoter sequence was prepared by in vitro transcription with a T7 Transcription Kit (Roche Diagnostics GmbH, Mannheim, Germany), according to the manufacturer’s instructions.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Incubation, Polymerase Chain Reaction, Expressing

    RacR binds to the ggt promoter region as shown by electrophoretic mobility shift assays (EMSA) . DIG-labeled PCR fragments (~50 fmol) containing the ggt, phoX , or Cj0200c promoter regions were incubated with RacR as indicated. (A) Influence of the phosphorylation of RacR on the binding to the ggt promoter. RacR was phosphorylated by RacScyto in the presence of ATP. (B) EMSA of the ggt, phoX , and Cj0200c promoter regions with phosphorylated RacR protein. The phoX and Cj0200c promoter regions were used as negative controls. RacR-P, phosphorylated RacR.

    Journal: Frontiers in Microbiology

    Article Title: The Campylobacter jejuni RacRS two-component system activates the glutamate synthesis by directly upregulating γ-glutamyltranspeptidase (GGT)

    doi: 10.3389/fmicb.2015.00567

    Figure Lengend Snippet: RacR binds to the ggt promoter region as shown by electrophoretic mobility shift assays (EMSA) . DIG-labeled PCR fragments (~50 fmol) containing the ggt, phoX , or Cj0200c promoter regions were incubated with RacR as indicated. (A) Influence of the phosphorylation of RacR on the binding to the ggt promoter. RacR was phosphorylated by RacScyto in the presence of ATP. (B) EMSA of the ggt, phoX , and Cj0200c promoter regions with phosphorylated RacR protein. The phoX and Cj0200c promoter regions were used as negative controls. RacR-P, phosphorylated RacR.

    Article Snippet: After electrophoresis the DNA was blotted on a hybond-N+ membrane (Amersham) and PCR fragments were visualized using α-DIG-AP, Fab fragments, and CSPD substrate (Roche).

    Techniques: Electrophoretic Mobility Shift Assay, Labeling, Polymerase Chain Reaction, Incubation, Binding Assay

    RacR binds to a specific region on the ggt promoter. (A) Nucleotide sequence and features of the ggt promoter. The start codon ATG is indicated in bold face, the putative -10 and -16 regions and ribosomal binding site (RBS) are underlined. A palindromic sequence is indicated with a horizontal bar. The previously identified RacR binding consensus sequence ( van der Stel et al., 2015 ) is indicated above the predicted RacR binding site, vertical lines indicate matching nucleotides. Arrows indicate the 5′ termini and direction of the primers used to generate the ggt promoter elements for the luciferase reporter plasmids and EMSA bait DNA. The transcriptional start site of the ggt gene identified by ( Dugar et al., 2013 ) is indicated with a hooked arrow. (B) Luciferase activities using different lengths of the region upstream of the ggt gene are determined in wt and racRS ::Cm mutant bacteria. Cultures were grown until late-log phase at oxygen limiting conditions with the addition of nitrate. Data represents the mean and SE of three independent experiments. (C) EMSA experiments using the different ggt promoter elements. DIG-labeled PCR fragments (~50 fmol) were mixed with or without 50 pmol RacR and 25 pmol RacScyto in the presence of ATP. RacR-P phosphorylated RacR.

    Journal: Frontiers in Microbiology

    Article Title: The Campylobacter jejuni RacRS two-component system activates the glutamate synthesis by directly upregulating γ-glutamyltranspeptidase (GGT)

    doi: 10.3389/fmicb.2015.00567

    Figure Lengend Snippet: RacR binds to a specific region on the ggt promoter. (A) Nucleotide sequence and features of the ggt promoter. The start codon ATG is indicated in bold face, the putative -10 and -16 regions and ribosomal binding site (RBS) are underlined. A palindromic sequence is indicated with a horizontal bar. The previously identified RacR binding consensus sequence ( van der Stel et al., 2015 ) is indicated above the predicted RacR binding site, vertical lines indicate matching nucleotides. Arrows indicate the 5′ termini and direction of the primers used to generate the ggt promoter elements for the luciferase reporter plasmids and EMSA bait DNA. The transcriptional start site of the ggt gene identified by ( Dugar et al., 2013 ) is indicated with a hooked arrow. (B) Luciferase activities using different lengths of the region upstream of the ggt gene are determined in wt and racRS ::Cm mutant bacteria. Cultures were grown until late-log phase at oxygen limiting conditions with the addition of nitrate. Data represents the mean and SE of three independent experiments. (C) EMSA experiments using the different ggt promoter elements. DIG-labeled PCR fragments (~50 fmol) were mixed with or without 50 pmol RacR and 25 pmol RacScyto in the presence of ATP. RacR-P phosphorylated RacR.

    Article Snippet: After electrophoresis the DNA was blotted on a hybond-N+ membrane (Amersham) and PCR fragments were visualized using α-DIG-AP, Fab fragments, and CSPD substrate (Roche).

    Techniques: Sequencing, Binding Assay, Luciferase, Mutagenesis, Labeling, Polymerase Chain Reaction