t4 rna ligase Promega Search Results


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  • 99
    Thermo Fisher t4 ligase
    T4 Ligase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3817 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    New England Biolabs t4 ligase 2
    T4 Ligase 2, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Promega t4 ligase
    T4 Ligase, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 1775 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche t4 ligase
    T4 Ligase, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 677 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega t4 ligase buffer
    T4 Ligase Buffer, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa t4 ligase buffer
    T4 Ligase Buffer, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega t4 rna ligase
    T4 Rna Ligase, supplied by Promega, used in various techniques. Bioz Stars score: 98/100, based on 626 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    TaKaRa t4 rna ligase
    T4 Rna Ligase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 733 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    MACHEREY NAGEL nucelospin rna purification kit
    Nucelospin Rna Purification Kit, supplied by MACHEREY NAGEL, used in various techniques. Bioz Stars score: 79/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche dig rna labeling mix
    Dig Rna Labeling Mix, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 225 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    New England Biolabs t4 rna ligase
    Determination of 5′- and 3′-Ends of Haloarchaeal Transcripts (A) Overview of the method. A recently developed method [ 28 ] was used to determine 5′-ends and 3′-ends of haloarchaeal transcripts. The overview schematically shows the different steps of the protocol. Circularization of transcripts with <t>T4</t> RNA ligase is only possible if their 5′-ends are monophosphorylated, in contrast to the 5′- triphosphate that is present when they are newly synthesized. This led to the initial belief that only processed transcripts can be analyzed [ 28 ], but it turned out that the method is also ideally suited to characterize primary transcripts (compare Discussion ). (B) Sequence of a PCR product representing a transcript with only one specific 3′-end (number 12 in Table 2 ). The stop and start codons of the gene are boxed, and the ligation point of the 5′-end and the 3′-end are denoted by an arrow. (C) Sequence of a PCR product representing a transcript with several 3′-ends (number 7 in Table 2 ). Different signal intensities are indicated by lines and the ligation point of the 5′-end and two different 3′-ends are denoted by arrows. (D) Sequences of two clones after cloning the PCR product shown in C (number 7 in Table 2 ). The results of two sequencing reactions of independent clones are shown. In both cases, the stop codon and start codon of the gene are boxed, and the ligation point of the 5′-end and the 3′-end are denoted by an arrow. (E) Results after sequencing ten clones. The sequences of five different 3′-ends and their number of occurrence are shown. The 5′-end was found to be identical in all ten cases.
    T4 Rna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 2987 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Integrated DNA Technologies crispr rna crrna sequence
    Generation of Igsf1 loss-of-function mice using <t>CRISPR-Cas9.</t> (a) PCR amplification of the targeted region in exon 18 of Igsf1 in the founder female (CRISPR.2, lane 1) and five of her progeny (3.1.1 to 3.1.5, lanes 2 to 6). Arrows at the right indicate the wild-type (top) or Δ312 (bottom) alleles. (b) Genomic organization (top) and <t>RNA</t> splicing (bottom) around exons 17 to 20 of the wild-type (left) and Δ312 alleles (right). Exons are presented as boxes and are numbered. Intervening introns are shown as lines. The deleted parts of exon 18 and intron 18 in the Δ312 allele are shown with a line at the top of the wild-type schematic (left). The novel exon in the Δ312 allele, which is derived from the 5′ end of exon 18, the 3′ end of intron 18 (labeled 18i), and all of exon 19, is shown with broken lines (right). Dark boxes denote translated sequences, whereas the pale boxes reflect untranslated regions. Genomic structure 5′ of exon 17 is not pictured. WT, wild-type.
    Crispr Rna Crrna Sequence, supplied by Integrated DNA Technologies, used in various techniques. Bioz Stars score: 80/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega t7 ribomax express large scale rna production system
    Generation of Igsf1 loss-of-function mice using <t>CRISPR-Cas9.</t> (a) PCR amplification of the targeted region in exon 18 of Igsf1 in the founder female (CRISPR.2, lane 1) and five of her progeny (3.1.1 to 3.1.5, lanes 2 to 6). Arrows at the right indicate the wild-type (top) or Δ312 (bottom) alleles. (b) Genomic organization (top) and <t>RNA</t> splicing (bottom) around exons 17 to 20 of the wild-type (left) and Δ312 alleles (right). Exons are presented as boxes and are numbered. Intervening introns are shown as lines. The deleted parts of exon 18 and intron 18 in the Δ312 allele are shown with a line at the top of the wild-type schematic (left). The novel exon in the Δ312 allele, which is derived from the 5′ end of exon 18, the 3′ end of intron 18 (labeled 18i), and all of exon 19, is shown with broken lines (right). Dark boxes denote translated sequences, whereas the pale boxes reflect untranslated regions. Genomic structure 5′ of exon 17 is not pictured. WT, wild-type.
    T7 Ribomax Express Large Scale Rna Production System, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1120 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    5 PRIME perfectpure cell tissue rna extraction kit
    Generation of Igsf1 loss-of-function mice using <t>CRISPR-Cas9.</t> (a) PCR amplification of the targeted region in exon 18 of Igsf1 in the founder female (CRISPR.2, lane 1) and five of her progeny (3.1.1 to 3.1.5, lanes 2 to 6). Arrows at the right indicate the wild-type (top) or Δ312 (bottom) alleles. (b) Genomic organization (top) and <t>RNA</t> splicing (bottom) around exons 17 to 20 of the wild-type (left) and Δ312 alleles (right). Exons are presented as boxes and are numbered. Intervening introns are shown as lines. The deleted parts of exon 18 and intron 18 in the Δ312 allele are shown with a line at the top of the wild-type schematic (left). The novel exon in the Δ312 allele, which is derived from the 5′ end of exon 18, the 3′ end of intron 18 (labeled 18i), and all of exon 19, is shown with broken lines (right). Dark boxes denote translated sequences, whereas the pale boxes reflect untranslated regions. Genomic structure 5′ of exon 17 is not pictured. WT, wild-type.
    Perfectpure Cell Tissue Rna Extraction Kit, supplied by 5 PRIME, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Promega t7 rna polymerase
    Generation of Igsf1 loss-of-function mice using <t>CRISPR-Cas9.</t> (a) PCR amplification of the targeted region in exon 18 of Igsf1 in the founder female (CRISPR.2, lane 1) and five of her progeny (3.1.1 to 3.1.5, lanes 2 to 6). Arrows at the right indicate the wild-type (top) or Δ312 (bottom) alleles. (b) Genomic organization (top) and <t>RNA</t> splicing (bottom) around exons 17 to 20 of the wild-type (left) and Δ312 alleles (right). Exons are presented as boxes and are numbered. Intervening introns are shown as lines. The deleted parts of exon 18 and intron 18 in the Δ312 allele are shown with a line at the top of the wild-type schematic (left). The novel exon in the Δ312 allele, which is derived from the 5′ end of exon 18, the 3′ end of intron 18 (labeled 18i), and all of exon 19, is shown with broken lines (right). Dark boxes denote translated sequences, whereas the pale boxes reflect untranslated regions. Genomic structure 5′ of exon 17 is not pictured. WT, wild-type.
    T7 Rna Polymerase, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 8588 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    New England Biolabs t7 rna polymerase
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    T7 Rna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 3044 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    GenePharma Company hairpin rnas shrnas
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    Hairpin Rnas Shrnas, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 99/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega rnase inhibitor
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    Rnase Inhibitor, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 8011 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega ligafast rapid dna ligation system
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    Ligafast Rapid Dna Ligation System, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 361 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega bamh i
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    Bamh I, supplied by Promega, used in various techniques. Bioz Stars score: 97/100, based on 209 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega rnase free dnase
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    Rnase Free Dnase, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 5992 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega miniprep dna purification systems
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    Miniprep Dna Purification Systems, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega celltiter blue viability assay
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    Celltiter Blue Viability Assay, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 175 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega collaborator plus transfast reagent
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    Collaborator Plus Transfast Reagent, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega rq1 dnase
    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of <t>T7</t> RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.
    Rq1 Dnase, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 8641 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Promega sv40 firefly luciferase
    Schema of the shPHD2 knockdown sites and the reporter constructs (A) Individual sequences of four small interfering RNA target sites against the PHD2 gene. (B) Schema of a classic hairpin carrying the site-2 sequence (shPHD2) and a control hairpin carrying the scramble sequence (shScramble). The H1 promoter drives the expression of a hairpin structure in which the sense and antisense strands of the small interfering RNA are connected by a 9-bp long loop sequence. In addition, a separate <t>5xHRE-SV40</t> promoter driving firefly luciferase (Fluc) is used to track shRNA activity in vitro and in vivo . 5x HRE, 5 repeat of hypoxia response elements; SV40, simian virus 40. (Reproduced from ref. 24 with permission).
    Sv40 Firefly Luciferase, supplied by Promega, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs spei
    Schema of the shPHD2 knockdown sites and the reporter constructs (A) Individual sequences of four small interfering RNA target sites against the PHD2 gene. (B) Schema of a classic hairpin carrying the site-2 sequence (shPHD2) and a control hairpin carrying the scramble sequence (shScramble). The H1 promoter drives the expression of a hairpin structure in which the sense and antisense strands of the small interfering RNA are connected by a 9-bp long loop sequence. In addition, a separate <t>5xHRE-SV40</t> promoter driving firefly luciferase (Fluc) is used to track shRNA activity in vitro and in vivo . 5x HRE, 5 repeat of hypoxia response elements; SV40, simian virus 40. (Reproduced from ref. 24 with permission).
    Spei, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 1936 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega pgem t easy vector
    Schema of the shPHD2 knockdown sites and the reporter constructs (A) Individual sequences of four small interfering RNA target sites against the PHD2 gene. (B) Schema of a classic hairpin carrying the site-2 sequence (shPHD2) and a control hairpin carrying the scramble sequence (shScramble). The H1 promoter drives the expression of a hairpin structure in which the sense and antisense strands of the small interfering RNA are connected by a 9-bp long loop sequence. In addition, a separate <t>5xHRE-SV40</t> promoter driving firefly luciferase (Fluc) is used to track shRNA activity in vitro and in vivo . 5x HRE, 5 repeat of hypoxia response elements; SV40, simian virus 40. (Reproduced from ref. 24 with permission).
    Pgem T Easy Vector, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 68436 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore mgcl2
    Schema of the shPHD2 knockdown sites and the reporter constructs (A) Individual sequences of four small interfering RNA target sites against the PHD2 gene. (B) Schema of a classic hairpin carrying the site-2 sequence (shPHD2) and a control hairpin carrying the scramble sequence (shScramble). The H1 promoter drives the expression of a hairpin structure in which the sense and antisense strands of the small interfering RNA are connected by a 9-bp long loop sequence. In addition, a separate <t>5xHRE-SV40</t> promoter driving firefly luciferase (Fluc) is used to track shRNA activity in vitro and in vivo . 5x HRE, 5 repeat of hypoxia response elements; SV40, simian virus 40. (Reproduced from ref. 24 with permission).
    Mgcl2, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 39277 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher cpg methyltransferase m sssi
    Schema of the shPHD2 knockdown sites and the reporter constructs (A) Individual sequences of four small interfering RNA target sites against the PHD2 gene. (B) Schema of a classic hairpin carrying the site-2 sequence (shPHD2) and a control hairpin carrying the scramble sequence (shScramble). The H1 promoter drives the expression of a hairpin structure in which the sense and antisense strands of the small interfering RNA are connected by a 9-bp long loop sequence. In addition, a separate <t>5xHRE-SV40</t> promoter driving firefly luciferase (Fluc) is used to track shRNA activity in vitro and in vivo . 5x HRE, 5 repeat of hypoxia response elements; SV40, simian virus 40. (Reproduced from ref. 24 with permission).
    Cpg Methyltransferase M Sssi, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Determination of 5′- and 3′-Ends of Haloarchaeal Transcripts (A) Overview of the method. A recently developed method [ 28 ] was used to determine 5′-ends and 3′-ends of haloarchaeal transcripts. The overview schematically shows the different steps of the protocol. Circularization of transcripts with T4 RNA ligase is only possible if their 5′-ends are monophosphorylated, in contrast to the 5′- triphosphate that is present when they are newly synthesized. This led to the initial belief that only processed transcripts can be analyzed [ 28 ], but it turned out that the method is also ideally suited to characterize primary transcripts (compare Discussion ). (B) Sequence of a PCR product representing a transcript with only one specific 3′-end (number 12 in Table 2 ). The stop and start codons of the gene are boxed, and the ligation point of the 5′-end and the 3′-end are denoted by an arrow. (C) Sequence of a PCR product representing a transcript with several 3′-ends (number 7 in Table 2 ). Different signal intensities are indicated by lines and the ligation point of the 5′-end and two different 3′-ends are denoted by arrows. (D) Sequences of two clones after cloning the PCR product shown in C (number 7 in Table 2 ). The results of two sequencing reactions of independent clones are shown. In both cases, the stop codon and start codon of the gene are boxed, and the ligation point of the 5′-end and the 3′-end are denoted by an arrow. (E) Results after sequencing ten clones. The sequences of five different 3′-ends and their number of occurrence are shown. The 5′-end was found to be identical in all ten cases.

    Journal: PLoS Genetics

    Article Title: Experimental Characterization of Cis-Acting Elements Important for Translation and Transcription in Halophilic Archaea

    doi: 10.1371/journal.pgen.0030229

    Figure Lengend Snippet: Determination of 5′- and 3′-Ends of Haloarchaeal Transcripts (A) Overview of the method. A recently developed method [ 28 ] was used to determine 5′-ends and 3′-ends of haloarchaeal transcripts. The overview schematically shows the different steps of the protocol. Circularization of transcripts with T4 RNA ligase is only possible if their 5′-ends are monophosphorylated, in contrast to the 5′- triphosphate that is present when they are newly synthesized. This led to the initial belief that only processed transcripts can be analyzed [ 28 ], but it turned out that the method is also ideally suited to characterize primary transcripts (compare Discussion ). (B) Sequence of a PCR product representing a transcript with only one specific 3′-end (number 12 in Table 2 ). The stop and start codons of the gene are boxed, and the ligation point of the 5′-end and the 3′-end are denoted by an arrow. (C) Sequence of a PCR product representing a transcript with several 3′-ends (number 7 in Table 2 ). Different signal intensities are indicated by lines and the ligation point of the 5′-end and two different 3′-ends are denoted by arrows. (D) Sequences of two clones after cloning the PCR product shown in C (number 7 in Table 2 ). The results of two sequencing reactions of independent clones are shown. In both cases, the stop codon and start codon of the gene are boxed, and the ligation point of the 5′-end and the 3′-end are denoted by an arrow. (E) Results after sequencing ten clones. The sequences of five different 3′-ends and their number of occurrence are shown. The 5′-end was found to be identical in all ten cases.

    Article Snippet: After denaturation at 65 °C for 10 min, total RNA was self-ligated by incubating 5–10 μg RNA with 40 U T4 RNA ligase (New England BioLabs), 10 U RNase Inhibitor (Promega), and 1 × T4 ligase buffer in a reaction volume of 25 μl at 37 °C for 1 h. After adjusting the total volume to 500 μl, proteins were removed by phenol chloroform extraction following standard protocols [ ].

    Techniques: Synthesized, Sequencing, Polymerase Chain Reaction, Ligation, Clone Assay

    Generation of Igsf1 loss-of-function mice using CRISPR-Cas9. (a) PCR amplification of the targeted region in exon 18 of Igsf1 in the founder female (CRISPR.2, lane 1) and five of her progeny (3.1.1 to 3.1.5, lanes 2 to 6). Arrows at the right indicate the wild-type (top) or Δ312 (bottom) alleles. (b) Genomic organization (top) and RNA splicing (bottom) around exons 17 to 20 of the wild-type (left) and Δ312 alleles (right). Exons are presented as boxes and are numbered. Intervening introns are shown as lines. The deleted parts of exon 18 and intron 18 in the Δ312 allele are shown with a line at the top of the wild-type schematic (left). The novel exon in the Δ312 allele, which is derived from the 5′ end of exon 18, the 3′ end of intron 18 (labeled 18i), and all of exon 19, is shown with broken lines (right). Dark boxes denote translated sequences, whereas the pale boxes reflect untranslated regions. Genomic structure 5′ of exon 17 is not pictured. WT, wild-type.

    Journal: Endocrinology

    Article Title: TRH Action Is Impaired in Pituitaries of Male IGSF1-Deficient Mice

    doi: 10.1210/en.2016-1788

    Figure Lengend Snippet: Generation of Igsf1 loss-of-function mice using CRISPR-Cas9. (a) PCR amplification of the targeted region in exon 18 of Igsf1 in the founder female (CRISPR.2, lane 1) and five of her progeny (3.1.1 to 3.1.5, lanes 2 to 6). Arrows at the right indicate the wild-type (top) or Δ312 (bottom) alleles. (b) Genomic organization (top) and RNA splicing (bottom) around exons 17 to 20 of the wild-type (left) and Δ312 alleles (right). Exons are presented as boxes and are numbered. Intervening introns are shown as lines. The deleted parts of exon 18 and intron 18 in the Δ312 allele are shown with a line at the top of the wild-type schematic (left). The novel exon in the Δ312 allele, which is derived from the 5′ end of exon 18, the 3′ end of intron 18 (labeled 18i), and all of exon 19, is shown with broken lines (right). Dark boxes denote translated sequences, whereas the pale boxes reflect untranslated regions. Genomic structure 5′ of exon 17 is not pictured. WT, wild-type.

    Article Snippet: DNA oligonucleotides encoding the CRISPR RNA (crRNA) sequence were purchased from Integrated DNA Technologies (Coralville, IA) ( ).

    Techniques: Mouse Assay, CRISPR, Polymerase Chain Reaction, Amplification, Derivative Assay, Labeling

    ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of T7 RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.

    Journal: Polymers

    Article Title: Size-Controllable Enzymatic Synthesis of Short Hairpin RNA Nanoparticles by Controlling the Rate of RNA Polymerization

    doi: 10.3390/polym10060589

    Figure Lengend Snippet: ( A ) Schematic illustration outlining the size-controlled synthesis of short hairpin RNA (shRNA) particles. By controlling concentrations of circular DNA in the rolling circle transcription (RCT) reaction at the same unit concentration of T7 RNA polymerase, the sizes of resulting RCT-mediated self-assembled shRNA particles could be controlled; ( B ) Scanning electron microscopy (SEM) images of shRNA particles showing their different sizes.

    Article Snippet: T7 RNA polymerase and ribonucleotide triphosphates (rNTPs) were purchased from New England BioLabs (NEB, Ipswich, MA, USA).

    Techniques: shRNA, Concentration Assay, Electron Microscopy

    Schema of the shPHD2 knockdown sites and the reporter constructs (A) Individual sequences of four small interfering RNA target sites against the PHD2 gene. (B) Schema of a classic hairpin carrying the site-2 sequence (shPHD2) and a control hairpin carrying the scramble sequence (shScramble). The H1 promoter drives the expression of a hairpin structure in which the sense and antisense strands of the small interfering RNA are connected by a 9-bp long loop sequence. In addition, a separate 5xHRE-SV40 promoter driving firefly luciferase (Fluc) is used to track shRNA activity in vitro and in vivo . 5x HRE, 5 repeat of hypoxia response elements; SV40, simian virus 40. (Reproduced from ref. 24 with permission).

    Journal: Methods in molecular biology (Clifton, N.J.)

    Article Title: Molecular Imaging of RNA Interference Therapy Targeting PHD2 for Treatment of Myocardial Ischemia

    doi: 10.1007/978-1-61737-982-6_13

    Figure Lengend Snippet: Schema of the shPHD2 knockdown sites and the reporter constructs (A) Individual sequences of four small interfering RNA target sites against the PHD2 gene. (B) Schema of a classic hairpin carrying the site-2 sequence (shPHD2) and a control hairpin carrying the scramble sequence (shScramble). The H1 promoter drives the expression of a hairpin structure in which the sense and antisense strands of the small interfering RNA are connected by a 9-bp long loop sequence. In addition, a separate 5xHRE-SV40 promoter driving firefly luciferase (Fluc) is used to track shRNA activity in vitro and in vivo . 5x HRE, 5 repeat of hypoxia response elements; SV40, simian virus 40. (Reproduced from ref. 24 with permission).

    Article Snippet: Plasmid pGL3 including SV40-Firefly luciferase (Promega).

    Techniques: Construct, Small Interfering RNA, Sequencing, Expressing, Luciferase, shRNA, Activity Assay, In Vitro, In Vivo