unr mrna sequence  (Thermo Fisher)


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    Structured Review

    Thermo Fisher unr mrna sequence
    SAABS assay. ( A ) SAABS procedure. A random 8mer oligodeoxynucleotide library (ROL) flanked by two PCR tags is first incubated with biotinylated <t>mRNA.</t> The mRNA bound ODN is then separated from free ODN by binding the biotinylated mRNA to a streptavidin coated Dynabead, which is then separated from the unbound sequence by a magnetic field. The bound sequence is then PCR amplified with S1 and CS2, restricted with NlaIII, concatenated by ligation, cloned in pZErO-1 and sequenced. ( B ) Frequency distribution of the antisense binding sites on the <t>unr</t> mRNA obtained from the SAABS assay. The 8mer sequences were retrieved from the sequenced clones and aligned with the mRNA sequence. Some of the sites identified correspond to sites found by the RT-ROL assay (13 and 46), whereas others were uniquely detected by the SAABS assay and denoted with an S prefix (S1, S3, S5 and S7).
    Unr Mrna Sequence, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/unr mrna sequence/product/Thermo Fisher
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    unr mrna sequence - by Bioz Stars, 2020-05
    85/100 stars

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    Images

    1) Product Images from "Identification and characterization of high affinity antisense PNAs for the human unr (upstream of N-ras) mRNA which is uniquely overexpressed in MCF-7 breast cancer cells"

    Article Title: Identification and characterization of high affinity antisense PNAs for the human unr (upstream of N-ras) mRNA which is uniquely overexpressed in MCF-7 breast cancer cells

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gki968

    SAABS assay. ( A ) SAABS procedure. A random 8mer oligodeoxynucleotide library (ROL) flanked by two PCR tags is first incubated with biotinylated mRNA. The mRNA bound ODN is then separated from free ODN by binding the biotinylated mRNA to a streptavidin coated Dynabead, which is then separated from the unbound sequence by a magnetic field. The bound sequence is then PCR amplified with S1 and CS2, restricted with NlaIII, concatenated by ligation, cloned in pZErO-1 and sequenced. ( B ) Frequency distribution of the antisense binding sites on the unr mRNA obtained from the SAABS assay. The 8mer sequences were retrieved from the sequenced clones and aligned with the mRNA sequence. Some of the sites identified correspond to sites found by the RT-ROL assay (13 and 46), whereas others were uniquely detected by the SAABS assay and denoted with an S prefix (S1, S3, S5 and S7).
    Figure Legend Snippet: SAABS assay. ( A ) SAABS procedure. A random 8mer oligodeoxynucleotide library (ROL) flanked by two PCR tags is first incubated with biotinylated mRNA. The mRNA bound ODN is then separated from free ODN by binding the biotinylated mRNA to a streptavidin coated Dynabead, which is then separated from the unbound sequence by a magnetic field. The bound sequence is then PCR amplified with S1 and CS2, restricted with NlaIII, concatenated by ligation, cloned in pZErO-1 and sequenced. ( B ) Frequency distribution of the antisense binding sites on the unr mRNA obtained from the SAABS assay. The 8mer sequences were retrieved from the sequenced clones and aligned with the mRNA sequence. Some of the sites identified correspond to sites found by the RT-ROL assay (13 and 46), whereas others were uniquely detected by the SAABS assay and denoted with an S prefix (S1, S3, S5 and S7).

    Techniques Used: Polymerase Chain Reaction, Incubation, Binding Assay, Sequencing, Amplification, Ligation, Clone Assay

    Dynabead-based dot blot assay to determine relative binding affinity of ODNs. ( A ) Determining the loading capacity of the streptavidin coated Dynabead by titrating 20 µl of bead solution in 40 µl total volume of 0.5 M NaCl with biotinylated radiolabeled unr mRNA. ( B ) Determining the µl of Dynabead bound RNA needed to completely bind 1 pmol of ODN5 in a total volume of 40 µl. ( C and D ) Solutions of RNA were incubated with 1 pmol of ODN [1–54 from RT-ROL assay and 57–68 (S1-S12) from the SAABS assay] and then incubated with 10 µl of Dynabeads and spotted on Nylon membrane. (C) is a photograph of blot showing equal loading of beads. (D) is a radiogram showing relative amounts of retained ODN. ODNs corresponding to circled spots were further studied by quantitative methods.
    Figure Legend Snippet: Dynabead-based dot blot assay to determine relative binding affinity of ODNs. ( A ) Determining the loading capacity of the streptavidin coated Dynabead by titrating 20 µl of bead solution in 40 µl total volume of 0.5 M NaCl with biotinylated radiolabeled unr mRNA. ( B ) Determining the µl of Dynabead bound RNA needed to completely bind 1 pmol of ODN5 in a total volume of 40 µl. ( C and D ) Solutions of RNA were incubated with 1 pmol of ODN [1–54 from RT-ROL assay and 57–68 (S1-S12) from the SAABS assay] and then incubated with 10 µl of Dynabeads and spotted on Nylon membrane. (C) is a photograph of blot showing equal loading of beads. (D) is a radiogram showing relative amounts of retained ODN. ODNs corresponding to circled spots were further studied by quantitative methods.

    Techniques Used: Dot Blot, Binding Assay, Incubation

    Related Articles

    Sequencing:

    Article Title: Identification and characterization of high affinity antisense PNAs for the human unr (upstream of N-ras) mRNA which is uniquely overexpressed in MCF-7 breast cancer cells
    Article Snippet: .. General Image clone 5285557 containing the unr mRNA sequence (D1S155E, NM_007158.2, GI: 20070240 and ATTC clone #7020864) was obtained from ATTC. pZErO-1 vector used for serial analysis of antisense binding sites (SAABS) analysis was from Invitrogen (Carlsbad, CA). .. Streptavidin coated Dynabeads were Dynabead M-280 Streptavidin from Dynal Biotech (Brown Deer, WI).

    Binding Assay:

    Article Title: Identification and characterization of high affinity antisense PNAs for the human unr (upstream of N-ras) mRNA which is uniquely overexpressed in MCF-7 breast cancer cells
    Article Snippet: .. General Image clone 5285557 containing the unr mRNA sequence (D1S155E, NM_007158.2, GI: 20070240 and ATTC clone #7020864) was obtained from ATTC. pZErO-1 vector used for serial analysis of antisense binding sites (SAABS) analysis was from Invitrogen (Carlsbad, CA). .. Streptavidin coated Dynabeads were Dynabead M-280 Streptavidin from Dynal Biotech (Brown Deer, WI).

    Plasmid Preparation:

    Article Title: Identification and characterization of high affinity antisense PNAs for the human unr (upstream of N-ras) mRNA which is uniquely overexpressed in MCF-7 breast cancer cells
    Article Snippet: .. General Image clone 5285557 containing the unr mRNA sequence (D1S155E, NM_007158.2, GI: 20070240 and ATTC clone #7020864) was obtained from ATTC. pZErO-1 vector used for serial analysis of antisense binding sites (SAABS) analysis was from Invitrogen (Carlsbad, CA). .. Streptavidin coated Dynabeads were Dynabead M-280 Streptavidin from Dynal Biotech (Brown Deer, WI).

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    Thermo Fisher csde1 mrna
    Csde1 Mrna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/csde1 mrna/product/Thermo Fisher
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