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g418  (Thermo Fisher)


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    Structured Review

    Thermo Fisher g418
    Stability of sfl genomes during continuous cell culture. (A) Six different cell lines were continuously cultured in the presence of <t>G418.</t> At the indicated time points (days [d]), cells were lysed, total RNA was prepared, and 4 μg was analyzed by Northern blotting. For comparison, given amounts of RNA containing subgenomic replicon transcribed in vitro were analyzed in parallel. To allow detection of sequence variants and of smaller HCV RNA pieces, the entire blot was hybridized under low-stringency conditions. (B) Genetic drift of the sfl genome in cell line 21-5. Cells were continuously cultured, and after 57 passages (corresponding to ∼6 months), total RNA was prepared. The coding region of the HCV polyprotein was amplified by long-distance RT-PCR in two overlapping fragments shown as bars below the ORF. For each fragment, two independent clones were sequenced. The boundary of the sequenced clones is indicated by a dashed vertical line. Amino acid substitutions are drawn as vertical lines at their respective positions within the HCV polyprotein. The hypervariable region 1 (HVR1) present at the N terminus of E2 is depicted as a shaded area. Six mutations were conserved between both sequenced clones, and they are indicated (stars).
    G418, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cell+transcriptome+sequencing+analysis/pmc00136109-62-9-11?v=Thermo+Fisher
    Average 99 stars, based on 1 article reviews
    g418 - by Bioz Stars, 2026-08
    99/100 stars

    Images

    1) Product Images from "Persistent and Transient Replication of Full-Length Hepatitis C Virus Genomes in Cell Culture"

    Article Title: Persistent and Transient Replication of Full-Length Hepatitis C Virus Genomes in Cell Culture

    Journal:

    doi: 10.1128/JVI.76.8.4008-4021.2002

    Stability of sfl genomes during continuous cell culture. (A) Six different cell lines were continuously cultured in the presence of G418. At the indicated time points (days [d]), cells were lysed, total RNA was prepared, and 4 μg was analyzed by Northern blotting. For comparison, given amounts of RNA containing subgenomic replicon transcribed in vitro were analyzed in parallel. To allow detection of sequence variants and of smaller HCV RNA pieces, the entire blot was hybridized under low-stringency conditions. (B) Genetic drift of the sfl genome in cell line 21-5. Cells were continuously cultured, and after 57 passages (corresponding to ∼6 months), total RNA was prepared. The coding region of the HCV polyprotein was amplified by long-distance RT-PCR in two overlapping fragments shown as bars below the ORF. For each fragment, two independent clones were sequenced. The boundary of the sequenced clones is indicated by a dashed vertical line. Amino acid substitutions are drawn as vertical lines at their respective positions within the HCV polyprotein. The hypervariable region 1 (HVR1) present at the N terminus of E2 is depicted as a shaded area. Six mutations were conserved between both sequenced clones, and they are indicated (stars).
    Figure Legend Snippet: Stability of sfl genomes during continuous cell culture. (A) Six different cell lines were continuously cultured in the presence of G418. At the indicated time points (days [d]), cells were lysed, total RNA was prepared, and 4 μg was analyzed by Northern blotting. For comparison, given amounts of RNA containing subgenomic replicon transcribed in vitro were analyzed in parallel. To allow detection of sequence variants and of smaller HCV RNA pieces, the entire blot was hybridized under low-stringency conditions. (B) Genetic drift of the sfl genome in cell line 21-5. Cells were continuously cultured, and after 57 passages (corresponding to ∼6 months), total RNA was prepared. The coding region of the HCV polyprotein was amplified by long-distance RT-PCR in two overlapping fragments shown as bars below the ORF. For each fragment, two independent clones were sequenced. The boundary of the sequenced clones is indicated by a dashed vertical line. Amino acid substitutions are drawn as vertical lines at their respective positions within the HCV polyprotein. The hypervariable region 1 (HVR1) present at the N terminus of E2 is depicted as a shaded area. Six mutations were conserved between both sequenced clones, and they are indicated (stars).

    Techniques Used: Cell Culture, Northern Blot, In Vitro, Sequencing, Amplification, Reverse Transcription Polymerase Chain Reaction, Clone Assay



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