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  • 99
    New England Biolabs t4 pnk
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    Processing of accumulated pre-rRNA in Atnuc-L1 mutant plants is accurate. A) Northern blot analysis using total RNA isolated from WT and Atnuc-L1-1 mutant plants and [γ 32P ] 5′-end labeled primers p34, p35, p36 and p41 to detect 5′ETS1 (lanes 1–3), 5′ETS2 (lanes 4–6), 18S (lanes 7–9) and 3′ETS (lanes 10–12) pre-rRNA sequences respectively. The asterisk and vertical bar indicate expected 5′ETS cleave off and exonucleolityc products (See also Figure S4 ). B) RNAseA/T1 protection analysis was carried out with a radiolabelled probe complementary to the 3′ETS (right). The assay was performed with total RNA from WT (lane 4) and Atnuc-L1 (lanes 5 and 6) or with yeast tRNA as a control (lane 3). A control lane loaded with undigested riboprobe is shown (lane 1). Lane 2, pBR322 digested with HpaII and 5′end labeled with <t>T4</t> PNK and [γ 32P ] ATP. C) Immunolocalization of fibrillarin in roots from WT and Atnuc-L1-1. Panel mFIB; Fibrillarin appears more abundant in the nucleolus of WT (a) than in the disorganized nucleolus of Atnuc-L1 plants (c) [18] . The nucleolar localization of fibrillarin practically overlaps the localization of AtNUC-L1 (b). Fibrillarin was detected with antibodies against mouse fibrillarin (mFIB 72B9) and Alexa-546 and AtNUC-L1 with antibodies against peptide AtNUC-L1 and Alexa-488. Chromatin in Atnuc-L1-1 is counterstained with DAPI (d). Bar, 10 µm.
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    Processing of accumulated pre-rRNA in Atnuc-L1 mutant plants is accurate. A) Northern blot analysis using total RNA isolated from WT and Atnuc-L1-1 mutant plants and [γ 32P ] 5′-end labeled primers p34, p35, p36 and p41 to detect 5′ETS1 (lanes 1–3), 5′ETS2 (lanes 4–6), 18S (lanes 7–9) and 3′ETS (lanes 10–12) pre-rRNA sequences respectively. The asterisk and vertical bar indicate expected 5′ETS cleave off and exonucleolityc products (See also Figure S4 ). B) RNAseA/T1 protection analysis was carried out with a radiolabelled probe complementary to the 3′ETS (right). The assay was performed with total RNA from WT (lane 4) and Atnuc-L1 (lanes 5 and 6) or with yeast tRNA as a control (lane 3). A control lane loaded with undigested riboprobe is shown (lane 1). Lane 2, pBR322 digested with HpaII and 5′end labeled with <t>T4</t> PNK and [γ 32P ] ATP. C) Immunolocalization of fibrillarin in roots from WT and Atnuc-L1-1. Panel mFIB; Fibrillarin appears more abundant in the nucleolus of WT (a) than in the disorganized nucleolus of Atnuc-L1 plants (c) [18] . The nucleolar localization of fibrillarin practically overlaps the localization of AtNUC-L1 (b). Fibrillarin was detected with antibodies against mouse fibrillarin (mFIB 72B9) and Alexa-546 and AtNUC-L1 with antibodies against peptide AtNUC-L1 and Alexa-488. Chromatin in Atnuc-L1-1 is counterstained with DAPI (d). Bar, 10 µm.
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    Enzymatics t4 pnk
    Processing of accumulated pre-rRNA in Atnuc-L1 mutant plants is accurate. A) Northern blot analysis using total RNA isolated from WT and Atnuc-L1-1 mutant plants and [γ 32P ] 5′-end labeled primers p34, p35, p36 and p41 to detect 5′ETS1 (lanes 1–3), 5′ETS2 (lanes 4–6), 18S (lanes 7–9) and 3′ETS (lanes 10–12) pre-rRNA sequences respectively. The asterisk and vertical bar indicate expected 5′ETS cleave off and exonucleolityc products (See also Figure S4 ). B) RNAseA/T1 protection analysis was carried out with a radiolabelled probe complementary to the 3′ETS (right). The assay was performed with total RNA from WT (lane 4) and Atnuc-L1 (lanes 5 and 6) or with yeast tRNA as a control (lane 3). A control lane loaded with undigested riboprobe is shown (lane 1). Lane 2, pBR322 digested with HpaII and 5′end labeled with <t>T4</t> PNK and [γ 32P ] ATP. C) Immunolocalization of fibrillarin in roots from WT and Atnuc-L1-1. Panel mFIB; Fibrillarin appears more abundant in the nucleolus of WT (a) than in the disorganized nucleolus of Atnuc-L1 plants (c) [18] . The nucleolar localization of fibrillarin practically overlaps the localization of AtNUC-L1 (b). Fibrillarin was detected with antibodies against mouse fibrillarin (mFIB 72B9) and Alexa-546 and AtNUC-L1 with antibodies against peptide AtNUC-L1 and Alexa-488. Chromatin in Atnuc-L1-1 is counterstained with DAPI (d). Bar, 10 µm.
    T4 Pnk, supplied by Enzymatics, used in various techniques. Bioz Stars score: 92/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Processing of accumulated pre-rRNA in Atnuc-L1 mutant plants is accurate. A) Northern blot analysis using total RNA isolated from WT and Atnuc-L1-1 mutant plants and [γ 32P ] 5′-end labeled primers p34, p35, p36 and p41 to detect 5′ETS1 (lanes 1–3), 5′ETS2 (lanes 4–6), 18S (lanes 7–9) and 3′ETS (lanes 10–12) pre-rRNA sequences respectively. The asterisk and vertical bar indicate expected 5′ETS cleave off and exonucleolityc products (See also Figure S4 ). B) RNAseA/T1 protection analysis was carried out with a radiolabelled probe complementary to the 3′ETS (right). The assay was performed with total RNA from WT (lane 4) and Atnuc-L1 (lanes 5 and 6) or with yeast tRNA as a control (lane 3). A control lane loaded with undigested riboprobe is shown (lane 1). Lane 2, pBR322 digested with HpaII and 5′end labeled with <t>T4</t> PNK and [γ 32P ] ATP. C) Immunolocalization of fibrillarin in roots from WT and Atnuc-L1-1. Panel mFIB; Fibrillarin appears more abundant in the nucleolus of WT (a) than in the disorganized nucleolus of Atnuc-L1 plants (c) [18] . The nucleolar localization of fibrillarin practically overlaps the localization of AtNUC-L1 (b). Fibrillarin was detected with antibodies against mouse fibrillarin (mFIB 72B9) and Alexa-546 and AtNUC-L1 with antibodies against peptide AtNUC-L1 and Alexa-488. Chromatin in Atnuc-L1-1 is counterstained with DAPI (d). Bar, 10 µm.
    T4 Polynucleotide Kinase 10 U µl, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Processing of accumulated pre-rRNA in Atnuc-L1 mutant plants is accurate. A) Northern blot analysis using total RNA isolated from WT and Atnuc-L1-1 mutant plants and [γ 32P ] 5′-end labeled primers p34, p35, p36 and p41 to detect 5′ETS1 (lanes 1–3), 5′ETS2 (lanes 4–6), 18S (lanes 7–9) and 3′ETS (lanes 10–12) pre-rRNA sequences respectively. The asterisk and vertical bar indicate expected 5′ETS cleave off and exonucleolityc products (See also Figure S4 ). B) RNAseA/T1 protection analysis was carried out with a radiolabelled probe complementary to the 3′ETS (right). The assay was performed with total RNA from WT (lane 4) and Atnuc-L1 (lanes 5 and 6) or with yeast tRNA as a control (lane 3). A control lane loaded with undigested riboprobe is shown (lane 1). Lane 2, pBR322 digested with HpaII and 5′end labeled with <t>T4</t> PNK and [γ 32P ] ATP. C) Immunolocalization of fibrillarin in roots from WT and Atnuc-L1-1. Panel mFIB; Fibrillarin appears more abundant in the nucleolus of WT (a) than in the disorganized nucleolus of Atnuc-L1 plants (c) [18] . The nucleolar localization of fibrillarin practically overlaps the localization of AtNUC-L1 (b). Fibrillarin was detected with antibodies against mouse fibrillarin (mFIB 72B9) and Alexa-546 and AtNUC-L1 with antibodies against peptide AtNUC-L1 and Alexa-488. Chromatin in Atnuc-L1-1 is counterstained with DAPI (d). Bar, 10 µm.
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    New England Biolabs dna polymerase i klenow fragment
    Processing of accumulated pre-rRNA in Atnuc-L1 mutant plants is accurate. A) Northern blot analysis using total RNA isolated from WT and Atnuc-L1-1 mutant plants and [γ 32P ] 5′-end labeled primers p34, p35, p36 and p41 to detect 5′ETS1 (lanes 1–3), 5′ETS2 (lanes 4–6), 18S (lanes 7–9) and 3′ETS (lanes 10–12) pre-rRNA sequences respectively. The asterisk and vertical bar indicate expected 5′ETS cleave off and exonucleolityc products (See also Figure S4 ). B) RNAseA/T1 protection analysis was carried out with a radiolabelled probe complementary to the 3′ETS (right). The assay was performed with total RNA from WT (lane 4) and Atnuc-L1 (lanes 5 and 6) or with yeast tRNA as a control (lane 3). A control lane loaded with undigested riboprobe is shown (lane 1). Lane 2, pBR322 digested with HpaII and 5′end labeled with <t>T4</t> PNK and [γ 32P ] ATP. C) Immunolocalization of fibrillarin in roots from WT and Atnuc-L1-1. Panel mFIB; Fibrillarin appears more abundant in the nucleolus of WT (a) than in the disorganized nucleolus of Atnuc-L1 plants (c) [18] . The nucleolar localization of fibrillarin practically overlaps the localization of AtNUC-L1 (b). Fibrillarin was detected with antibodies against mouse fibrillarin (mFIB 72B9) and Alexa-546 and AtNUC-L1 with antibodies against peptide AtNUC-L1 and Alexa-488. Chromatin in Atnuc-L1-1 is counterstained with DAPI (d). Bar, 10 µm.
    Dna Polymerase I Klenow Fragment, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 842 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Processing of accumulated pre-rRNA in Atnuc-L1 mutant plants is accurate. A) Northern blot analysis using total RNA isolated from WT and Atnuc-L1-1 mutant plants and [γ 32P ] 5′-end labeled primers p34, p35, p36 and p41 to detect 5′ETS1 (lanes 1–3), 5′ETS2 (lanes 4–6), 18S (lanes 7–9) and 3′ETS (lanes 10–12) pre-rRNA sequences respectively. The asterisk and vertical bar indicate expected 5′ETS cleave off and exonucleolityc products (See also Figure S4 ). B) RNAseA/T1 protection analysis was carried out with a radiolabelled probe complementary to the 3′ETS (right). The assay was performed with total RNA from WT (lane 4) and Atnuc-L1 (lanes 5 and 6) or with yeast tRNA as a control (lane 3). A control lane loaded with undigested riboprobe is shown (lane 1). Lane 2, pBR322 digested with HpaII and 5′end labeled with T4 PNK and [γ 32P ] ATP. C) Immunolocalization of fibrillarin in roots from WT and Atnuc-L1-1. Panel mFIB; Fibrillarin appears more abundant in the nucleolus of WT (a) than in the disorganized nucleolus of Atnuc-L1 plants (c) [18] . The nucleolar localization of fibrillarin practically overlaps the localization of AtNUC-L1 (b). Fibrillarin was detected with antibodies against mouse fibrillarin (mFIB 72B9) and Alexa-546 and AtNUC-L1 with antibodies against peptide AtNUC-L1 and Alexa-488. Chromatin in Atnuc-L1-1 is counterstained with DAPI (d). Bar, 10 µm.

    Journal: PLoS Genetics

    Article Title: Nucleolin Is Required for DNA Methylation State and the Expression of rRNA Gene Variants in Arabidopsis thaliana

    doi: 10.1371/journal.pgen.1001225

    Figure Lengend Snippet: Processing of accumulated pre-rRNA in Atnuc-L1 mutant plants is accurate. A) Northern blot analysis using total RNA isolated from WT and Atnuc-L1-1 mutant plants and [γ 32P ] 5′-end labeled primers p34, p35, p36 and p41 to detect 5′ETS1 (lanes 1–3), 5′ETS2 (lanes 4–6), 18S (lanes 7–9) and 3′ETS (lanes 10–12) pre-rRNA sequences respectively. The asterisk and vertical bar indicate expected 5′ETS cleave off and exonucleolityc products (See also Figure S4 ). B) RNAseA/T1 protection analysis was carried out with a radiolabelled probe complementary to the 3′ETS (right). The assay was performed with total RNA from WT (lane 4) and Atnuc-L1 (lanes 5 and 6) or with yeast tRNA as a control (lane 3). A control lane loaded with undigested riboprobe is shown (lane 1). Lane 2, pBR322 digested with HpaII and 5′end labeled with T4 PNK and [γ 32P ] ATP. C) Immunolocalization of fibrillarin in roots from WT and Atnuc-L1-1. Panel mFIB; Fibrillarin appears more abundant in the nucleolus of WT (a) than in the disorganized nucleolus of Atnuc-L1 plants (c) [18] . The nucleolar localization of fibrillarin practically overlaps the localization of AtNUC-L1 (b). Fibrillarin was detected with antibodies against mouse fibrillarin (mFIB 72B9) and Alexa-546 and AtNUC-L1 with antibodies against peptide AtNUC-L1 and Alexa-488. Chromatin in Atnuc-L1-1 is counterstained with DAPI (d). Bar, 10 µm.

    Article Snippet: For detection of pre-rRNA precursors and small RNA, 10 pmoles of oligonucleotide primers were 5′end labeled using 50 µCi of [γ32 P] ATP (6000 Ci/mmol) and T4 PNK (Promega).

    Techniques: Mutagenesis, Northern Blot, Isolation, Labeling