human cot1 dna  (Thermo Fisher)


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    Name:
    Human Cot 1 DNA
    Description:
    Human Cot 1 DNA is commonly used to block nonspecific hybridization in microarray screening It can also be used to suppress repetitive DNA sequences for the direct mapping of human DNA or mapping genomic clones to panels of somatic cell hybrids for chromosome localization by Southern blotting Human Cot 1 DNA is effective as a library screening probe for somatic cell hybrid libraries and flow sorted chromosome libraries made from somatic cell hybrids About Cot 1 DNA Human Cot 1 DNA is placental DNA that is predominantly 50 to 300 bp in size and enriched for repetitive DNA sequences such as the Alu and Kpn family members The amount supplied per package is sufficient for 5 10 Southern or 500 in situ hybridizations Performance and quality testingPurity and DNA size are verified by agarose gel electrophoresis Concentration is verified spectrophotometrically by diluting Human Cot 1 DNA 1 100 in 50 mM NaOH and using the conversion factor 0 033 µg µL A260 Other methods used to determine concentration may yield varying results
    Catalog Number:
    15279011
    Price:
    None
    Applications:
    Cell Analysis|Cellular Imaging|ChIP-on-Chip|Fluorescence In Situ Hybridization|Genotyping & Genomic Profiling|In Situ Hybridization (ISH)|RNAi, Epigenetics & Non-Coding RNA Research|Chromatin Biology|Gene Expression Analysis & Genotyping|Array CGH|Microarray Hybridization & General Reagents
    Category:
    Lab Reagents and Chemicals
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    Structured Review

    Thermo Fisher human cot1 dna
    The human Xi is partially reactivated upon cell fusion-mediated reprogramming. ( a ) Expression of human Xi transcripts examined by allele-specific RNA-seq in female human fibroblasts (clone b1) before (day 0) and after (day 5/6) mESC-fusion. Data points show Xi expression of 113 genes along the X chromosome ( x axis) where genes that escape XCI before reprogramming (blue closed circles),, or that are sensitive to reactivation following fusion (day 5 green open circles; day 6 green closed circles) are highlighted. Xi expression is represented as the ratio of reads overlapping the minor (Xi) allele versus the total of both alleles (Xi+Xa) and result from two biological replicates. Genes were classified accordingly to Xi expression ratio and significance 62 , as detailed in the Methods. ( b ) RNA-FISH analyses of candidate reactivation-refractory ( HDAC8 ) and reactivation-sensitive ( RP11-706O15.1 , WWC3 ) genes in human fibroblasts before (day 0) and after mESC-fusion (day 5). Histogram plots show the percentage of human nuclei (heterokaryon/hybrid cells; n = 44/83, 33/74, 48/81 for HDAC8 , RP11-706O15.1 and WWC3 , respectively) with one, two, or > 2 punctate RNA signals. Representative confocal images showing mono-allelic expression of WWC3 (red) in human nuclei before (day 0) and bi-allelic expression after (day 5) mESC-fusion (arrowed) where mouse nuclei were discriminated using mCotI probe (green). Arrows indicate transcribed loci (that is, punctate RNA signals). Scale bar, 5 μm. ( c ) Confocal images of hFxmESC heterokaryons where either one (top) or two (lower) transcribed X chromosomes (arrowed) were evident. Mouse <t>Cot1</t> probe identifies mouse (ESC-derived) nuclei. X chr RNA paints were pre-treated with human Cot1 <t>DNA</t> to diminish the detection of repeat-rich elements 63 within the transcribed X chromosome domains. Xa* marks the smaller (and Xa the larger) domain. Scale bar, 5 μm. ( d ) Box plots showing the volume of the RNA (top) and DNA (bottom) X domains detected by sequential RNA/DNA FISH in hFxmESC expressing one (single domain) or two (two domains) X chromosomes (shown in Supplementary Fig. 5b ). Significant volume differences are highlighted ( P values) and were estimated by Wilcoxon signed-rank test or Mann–Whitney test, for the comparison of Xa/Xa* and Xa/Xi, respectively. Data represent at least 50 cells in each graph. See also Supplementary Fig. 5 .
    Human Cot 1 DNA is commonly used to block nonspecific hybridization in microarray screening It can also be used to suppress repetitive DNA sequences for the direct mapping of human DNA or mapping genomic clones to panels of somatic cell hybrids for chromosome localization by Southern blotting Human Cot 1 DNA is effective as a library screening probe for somatic cell hybrid libraries and flow sorted chromosome libraries made from somatic cell hybrids About Cot 1 DNA Human Cot 1 DNA is placental DNA that is predominantly 50 to 300 bp in size and enriched for repetitive DNA sequences such as the Alu and Kpn family members The amount supplied per package is sufficient for 5 10 Southern or 500 in situ hybridizations Performance and quality testingPurity and DNA size are verified by agarose gel electrophoresis Concentration is verified spectrophotometrically by diluting Human Cot 1 DNA 1 100 in 50 mM NaOH and using the conversion factor 0 033 µg µL A260 Other methods used to determine concentration may yield varying results
    https://www.bioz.com/result/human cot1 dna/product/Thermo Fisher
    Average 99 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    human cot1 dna - by Bioz Stars, 2020-09
    99/100 stars

    Images

    1) Product Images from "Ordered chromatin changes and human X chromosome reactivation by cell fusion-mediated pluripotent reprogramming"

    Article Title: Ordered chromatin changes and human X chromosome reactivation by cell fusion-mediated pluripotent reprogramming

    Journal: Nature Communications

    doi: 10.1038/ncomms12354

    The human Xi is partially reactivated upon cell fusion-mediated reprogramming. ( a ) Expression of human Xi transcripts examined by allele-specific RNA-seq in female human fibroblasts (clone b1) before (day 0) and after (day 5/6) mESC-fusion. Data points show Xi expression of 113 genes along the X chromosome ( x axis) where genes that escape XCI before reprogramming (blue closed circles),, or that are sensitive to reactivation following fusion (day 5 green open circles; day 6 green closed circles) are highlighted. Xi expression is represented as the ratio of reads overlapping the minor (Xi) allele versus the total of both alleles (Xi+Xa) and result from two biological replicates. Genes were classified accordingly to Xi expression ratio and significance 62 , as detailed in the Methods. ( b ) RNA-FISH analyses of candidate reactivation-refractory ( HDAC8 ) and reactivation-sensitive ( RP11-706O15.1 , WWC3 ) genes in human fibroblasts before (day 0) and after mESC-fusion (day 5). Histogram plots show the percentage of human nuclei (heterokaryon/hybrid cells; n = 44/83, 33/74, 48/81 for HDAC8 , RP11-706O15.1 and WWC3 , respectively) with one, two, or > 2 punctate RNA signals. Representative confocal images showing mono-allelic expression of WWC3 (red) in human nuclei before (day 0) and bi-allelic expression after (day 5) mESC-fusion (arrowed) where mouse nuclei were discriminated using mCotI probe (green). Arrows indicate transcribed loci (that is, punctate RNA signals). Scale bar, 5 μm. ( c ) Confocal images of hFxmESC heterokaryons where either one (top) or two (lower) transcribed X chromosomes (arrowed) were evident. Mouse Cot1 probe identifies mouse (ESC-derived) nuclei. X chr RNA paints were pre-treated with human Cot1 DNA to diminish the detection of repeat-rich elements 63 within the transcribed X chromosome domains. Xa* marks the smaller (and Xa the larger) domain. Scale bar, 5 μm. ( d ) Box plots showing the volume of the RNA (top) and DNA (bottom) X domains detected by sequential RNA/DNA FISH in hFxmESC expressing one (single domain) or two (two domains) X chromosomes (shown in Supplementary Fig. 5b ). Significant volume differences are highlighted ( P values) and were estimated by Wilcoxon signed-rank test or Mann–Whitney test, for the comparison of Xa/Xa* and Xa/Xi, respectively. Data represent at least 50 cells in each graph. See also Supplementary Fig. 5 .
    Figure Legend Snippet: The human Xi is partially reactivated upon cell fusion-mediated reprogramming. ( a ) Expression of human Xi transcripts examined by allele-specific RNA-seq in female human fibroblasts (clone b1) before (day 0) and after (day 5/6) mESC-fusion. Data points show Xi expression of 113 genes along the X chromosome ( x axis) where genes that escape XCI before reprogramming (blue closed circles),, or that are sensitive to reactivation following fusion (day 5 green open circles; day 6 green closed circles) are highlighted. Xi expression is represented as the ratio of reads overlapping the minor (Xi) allele versus the total of both alleles (Xi+Xa) and result from two biological replicates. Genes were classified accordingly to Xi expression ratio and significance 62 , as detailed in the Methods. ( b ) RNA-FISH analyses of candidate reactivation-refractory ( HDAC8 ) and reactivation-sensitive ( RP11-706O15.1 , WWC3 ) genes in human fibroblasts before (day 0) and after mESC-fusion (day 5). Histogram plots show the percentage of human nuclei (heterokaryon/hybrid cells; n = 44/83, 33/74, 48/81 for HDAC8 , RP11-706O15.1 and WWC3 , respectively) with one, two, or > 2 punctate RNA signals. Representative confocal images showing mono-allelic expression of WWC3 (red) in human nuclei before (day 0) and bi-allelic expression after (day 5) mESC-fusion (arrowed) where mouse nuclei were discriminated using mCotI probe (green). Arrows indicate transcribed loci (that is, punctate RNA signals). Scale bar, 5 μm. ( c ) Confocal images of hFxmESC heterokaryons where either one (top) or two (lower) transcribed X chromosomes (arrowed) were evident. Mouse Cot1 probe identifies mouse (ESC-derived) nuclei. X chr RNA paints were pre-treated with human Cot1 DNA to diminish the detection of repeat-rich elements 63 within the transcribed X chromosome domains. Xa* marks the smaller (and Xa the larger) domain. Scale bar, 5 μm. ( d ) Box plots showing the volume of the RNA (top) and DNA (bottom) X domains detected by sequential RNA/DNA FISH in hFxmESC expressing one (single domain) or two (two domains) X chromosomes (shown in Supplementary Fig. 5b ). Significant volume differences are highlighted ( P values) and were estimated by Wilcoxon signed-rank test or Mann–Whitney test, for the comparison of Xa/Xa* and Xa/Xi, respectively. Data represent at least 50 cells in each graph. See also Supplementary Fig. 5 .

    Techniques Used: Expressing, RNA Sequencing Assay, Fluorescence In Situ Hybridization, Derivative Assay, MANN-WHITNEY

    Related Articles

    Fluorescence:

    Article Title: Misbehaviour of XIST RNA in Breast Cancer Cells
    Article Snippet: .. Cot-1 probe was obtained labeling 100 ng of Cot-1 DNA (1 mg/ml, Invitrogen) by Prime-It Fluor, Fluorescence labeling Kit with FITC-dUTP. .. The probes were then ethanol precipitated, washed in 70% ethanol, air dried and resuspended in 15 µL of hybridization buffer (50% formamide, 4× SSC, 20% dextran sulfate, 40 mM VRC, 0,4% BSA).

    Labeling:

    Article Title: Methods for high throughput validation of amplified fragment pools of BAC DNA for constructing high resolution CGH arrays
    Article Snippet: .. The labeled probes were precipitated in ethanol with (or without) 50 μg Cot-1 DNA (Invitrogen) and redissolved in 15 μl of hybridization solution (50% formamide, 2X SSC, 10% dextran sulfate, 4% SDS). ..

    Article Title: Misbehaviour of XIST RNA in Breast Cancer Cells
    Article Snippet: .. Cot-1 probe was obtained labeling 100 ng of Cot-1 DNA (1 mg/ml, Invitrogen) by Prime-It Fluor, Fluorescence labeling Kit with FITC-dUTP. .. The probes were then ethanol precipitated, washed in 70% ethanol, air dried and resuspended in 15 µL of hybridization buffer (50% formamide, 4× SSC, 20% dextran sulfate, 40 mM VRC, 0,4% BSA).

    Evaporation:

    Article Title: Cross-species hybridisation of human and bovine orthologous genes on high density cDNA microarrays
    Article Snippet: .. After concentrating cDNAs by evaporation in a SpeedVac, labelled targets were resuspended in 20 μl of hybridisation buffer (10 μg polydA and 20 μg Human Cot1 DNA,-Invitrogen; DIG-Easy Hybridisation mix -Roche). .. After thorough resuspension, the cDNA was denatured by heating at 95° C for 5 mins followed by 20 mins. at 42° C to enable annealing of the blocking reagents to repetitive sequences within the target cDNAs.

    BAC Assay:

    Article Title: Reversible immortalisation enables genetic correction of human muscle progenitors and engineering of next‐generation human artificial chromosomes for Duchenne muscular dystrophy
    Article Snippet: .. Digoxigenin‐labelled (Roche) human COT‐1 DNA (Invitrogen) and biotin‐labelled BAC DNA RP11‐954B16 (located in the Dystrophin genomic region, Children's Hospital Oakland Research Institute) were used for the detection of DYS‐HAC2 in DT40 (DYS‐HAC2), A9(DYS‐HAC2)‐9 and CHO(DYS‐HAC2)‐7 cells. .. Digoxigenin‐labelled p11‐4 human alpha satellite (centromeres of chromosome 13 and 21, hChr 13/21(cen)) and biotin‐labelled RP11‐954B16 were used for the detection of DYS‐HAC2 in riDMD(DYS‐HAC2) clones.

    Polymerase Chain Reaction:

    Article Title: A Novel Approach for Determining Cancer Genomic Breakpoints in the Presence of Normal DNA
    Article Snippet: .. The PCR products were purified with DNA Clean-up and Concentrator-5 (Zymo Research, Orange, CA), resuspended in 3×SSC and printed on poly-L-lysine slides at 0.1 mg/ml along with Human Cot-1 DNA (Invitrogen, Carlsbad, CA), which is enriched for repetitive sequences, and herring sperm DNA (Promega, Madison, WI), which was used as nonspecific control. .. The printing procedure has been described and essentially followed the manual of the DeRisi arrayer with silicon microcontact printing pins (Parallel Synthesis Technologies, Inc. Santa Clara, CA) , .

    Purification:

    Article Title: A Novel Approach for Determining Cancer Genomic Breakpoints in the Presence of Normal DNA
    Article Snippet: .. The PCR products were purified with DNA Clean-up and Concentrator-5 (Zymo Research, Orange, CA), resuspended in 3×SSC and printed on poly-L-lysine slides at 0.1 mg/ml along with Human Cot-1 DNA (Invitrogen, Carlsbad, CA), which is enriched for repetitive sequences, and herring sperm DNA (Promega, Madison, WI), which was used as nonspecific control. .. The printing procedure has been described and essentially followed the manual of the DeRisi arrayer with silicon microcontact printing pins (Parallel Synthesis Technologies, Inc. Santa Clara, CA) , .

    Fluorescence In Situ Hybridization:

    Article Title: Use of a Human Artificial Chromosome for Delivering Trophic Factors in a Rodent Model of Amyotrophic Lateral Sclerosis
    Article Snippet: .. FISH analyses were performed using fixed metaphase spreads of each cell hybrid using digoxigenin-labeled (Roche, Basel, Switzerland) human COT-1 DNA (Invitrogen), digoxigenin-labeled p11-4 (hChr.21-derived alpha-satellite clone), and biotin-labeled PAC DNA, essentially as described previously. ..

    Hybridization:

    Article Title: Methods for high throughput validation of amplified fragment pools of BAC DNA for constructing high resolution CGH arrays
    Article Snippet: .. The labeled probes were precipitated in ethanol with (or without) 50 μg Cot-1 DNA (Invitrogen) and redissolved in 15 μl of hybridization solution (50% formamide, 2X SSC, 10% dextran sulfate, 4% SDS). ..

    Article Title: Cross-species hybridisation of human and bovine orthologous genes on high density cDNA microarrays
    Article Snippet: .. After concentrating cDNAs by evaporation in a SpeedVac, labelled targets were resuspended in 20 μl of hybridisation buffer (10 μg polydA and 20 μg Human Cot1 DNA,-Invitrogen; DIG-Easy Hybridisation mix -Roche). .. After thorough resuspension, the cDNA was denatured by heating at 95° C for 5 mins followed by 20 mins. at 42° C to enable annealing of the blocking reagents to repetitive sequences within the target cDNAs.

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  • 99
    Thermo Fisher human cot 1 dna
    Human Cot 1 Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 387 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/human cot 1 dna/product/Thermo Fisher
    Average 99 stars, based on 387 article reviews
    Price from $9.99 to $1999.99
    human cot 1 dna - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

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