dna extraction kit  (TaKaRa)


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    Name:
    DNA Blunting Kit
    Description:
    The DNA Blunting Kit allows the conversion of 3 and 5 overhangs to blunt or flush ends This conversion is accomplished simultaneously by the 3 →5 exonuclease and 5 →3 polymerase activities of T4 DNA Polymerase The resulting blunt ended DNA can then be ligated into a vector using the same optimized buffer system employed in our DNA Ligation Kit Mighty Mix TaKaRa DNA Ligation Kit LONG and Quick DNA ligation kit products The reaction can then be used directly in bacterial transformation or in vitro packaging procedures without the need for further DNA purification
    Catalog Number:
    6025
    Price:
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    Category:
    DNA Blunting Kit Ligation kits Cloning
    Size:
    20 Rxns
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    Structured Review

    TaKaRa dna extraction kit
    Detection of MEV in clinical samples by nanoPCR assay. Lane M: DL2000 <t>DNA</t> Maker (TaKaRa, <t>Dalian,</t> China); lane 1: MEV genome as template; lane 2: plasmid DNA as template; lanes 3: negtive control, lanes 4–21: DNA from clinical fecal samples as template.
    The DNA Blunting Kit allows the conversion of 3 and 5 overhangs to blunt or flush ends This conversion is accomplished simultaneously by the 3 →5 exonuclease and 5 →3 polymerase activities of T4 DNA Polymerase The resulting blunt ended DNA can then be ligated into a vector using the same optimized buffer system employed in our DNA Ligation Kit Mighty Mix TaKaRa DNA Ligation Kit LONG and Quick DNA ligation kit products The reaction can then be used directly in bacterial transformation or in vitro packaging procedures without the need for further DNA purification
    https://www.bioz.com/result/dna extraction kit/product/TaKaRa
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    dna extraction kit - by Bioz Stars, 2021-03
    99/100 stars

    Images

    1) Product Images from "Development of a nanoparticle-assisted PCR (nanoPCR) assay for detection of mink enteritis virus (MEV) and genetic characterization of the NS1 gene in four Chinese MEV strains"

    Article Title: Development of a nanoparticle-assisted PCR (nanoPCR) assay for detection of mink enteritis virus (MEV) and genetic characterization of the NS1 gene in four Chinese MEV strains

    Journal: BMC Veterinary Research

    doi: 10.1186/s12917-014-0312-6

    Detection of MEV in clinical samples by nanoPCR assay. Lane M: DL2000 DNA Maker (TaKaRa, Dalian, China); lane 1: MEV genome as template; lane 2: plasmid DNA as template; lanes 3: negtive control, lanes 4–21: DNA from clinical fecal samples as template.
    Figure Legend Snippet: Detection of MEV in clinical samples by nanoPCR assay. Lane M: DL2000 DNA Maker (TaKaRa, Dalian, China); lane 1: MEV genome as template; lane 2: plasmid DNA as template; lanes 3: negtive control, lanes 4–21: DNA from clinical fecal samples as template.

    Techniques Used: Nano PCR Assay, Plasmid Preparation

    2) Product Images from "Integrated DNA and RNA extraction using magnetic beads from viral pathogens causing acute respiratory infections"

    Article Title: Integrated DNA and RNA extraction using magnetic beads from viral pathogens causing acute respiratory infections

    Journal: Scientific Reports

    doi: 10.1038/srep45199

    The comparison of Ct values obtained from the different nucleic acid extraction methods. F1: RNA carrier A; F2:RNA carrier B; F3: no RNA carrier; F4: TIANGEN; F5: TaKaRa RNA/DNA extraction kits that extract RNA and DNA separately. For different viral nucleic acid extraction methods, For Ct values of RSV (RNA), there were statistical differences among all the groups through SNK test (F = 106.73, P = 0.00). For Ct values of ADV (DNA), the differences among groups F5, F4 and F1 were not statistically significant, but there were statistical differences among group F2, F3 and group (F1, F4, F5) (F = 75.14, P = 0.00).
    Figure Legend Snippet: The comparison of Ct values obtained from the different nucleic acid extraction methods. F1: RNA carrier A; F2:RNA carrier B; F3: no RNA carrier; F4: TIANGEN; F5: TaKaRa RNA/DNA extraction kits that extract RNA and DNA separately. For different viral nucleic acid extraction methods, For Ct values of RSV (RNA), there were statistical differences among all the groups through SNK test (F = 106.73, P = 0.00). For Ct values of ADV (DNA), the differences among groups F5, F4 and F1 were not statistically significant, but there were statistical differences among group F2, F3 and group (F1, F4, F5) (F = 75.14, P = 0.00).

    Techniques Used: DNA Extraction

    The Ct values of RNA (RSV) or DNA (ADV) obtained from the duplex RT-qPCR based on the assessment of different influencing factors. For GTC concentrations ( A ) for RSV, there were no statistical differences among groupA2, A3 and A4, but there were statistical differences among group A2, A3, A4 and group A1 (F = 51.29, P = 0.00); for ADV, the differences among all the groups weren’t statistically significant (F = 1.94, P = 0.20). For DTT concentrations ( B ) for RSV, there were no statistical differences among group B3, B4, B5 and B6, but there were statistical differences among group (B3, B4, B5, B6), group B2 and group B1 (F = 38.84, P = 0.00); for ADV, there were no statistical differences among group B3, B4, B5, B6, but there were statistical differences among group (B3, B4, B5, B6), group B2, group B1 (F = 10.18, P = 0.00). For magnetic bead amounts ( C ) for RSV, there were no statistical differences among group C2, C3 and C4, but there were statistical differences among group C2, C3, C4 and group C1 (F = 185.32, P = 0.00); for ADV, there were no statistical differences among group C2, C3 and C4, but there were statistical differences among group C2, C3, C4 and group C1 (F = 42.11, P = 0.00). For temperatures ( D ) for RSV, there were no statistical differences among group D2, D3 and D4, but there were statistical differences between group D2, D3, D4 and group D1 (F = 44.56, P = 0.00); for ADV, there were no statistical differences among group D2, D3 and D4, but there were statistical differences between group D2, D3, D4 and group D1 (F = 77.52, P = 0.00). For pH ( E ) for RSV, there was no statistical differences between group E5 and E6, and so was between group E3 and E4, but there were statistical differences among group (E5, E6), group (E3, E4), group E1 and group E2 (F = 85.36, P = 0.00); for ADV, there was no statistical differences between group E5 and E6, and so was between group E3 and E4, but there were statistical differences among group (E5, E6), group (E3, E4), group E1 and group E2 (F = 1370.60, P = 0.00).
    Figure Legend Snippet: The Ct values of RNA (RSV) or DNA (ADV) obtained from the duplex RT-qPCR based on the assessment of different influencing factors. For GTC concentrations ( A ) for RSV, there were no statistical differences among groupA2, A3 and A4, but there were statistical differences among group A2, A3, A4 and group A1 (F = 51.29, P = 0.00); for ADV, the differences among all the groups weren’t statistically significant (F = 1.94, P = 0.20). For DTT concentrations ( B ) for RSV, there were no statistical differences among group B3, B4, B5 and B6, but there were statistical differences among group (B3, B4, B5, B6), group B2 and group B1 (F = 38.84, P = 0.00); for ADV, there were no statistical differences among group B3, B4, B5, B6, but there were statistical differences among group (B3, B4, B5, B6), group B2, group B1 (F = 10.18, P = 0.00). For magnetic bead amounts ( C ) for RSV, there were no statistical differences among group C2, C3 and C4, but there were statistical differences among group C2, C3, C4 and group C1 (F = 185.32, P = 0.00); for ADV, there were no statistical differences among group C2, C3 and C4, but there were statistical differences among group C2, C3, C4 and group C1 (F = 42.11, P = 0.00). For temperatures ( D ) for RSV, there were no statistical differences among group D2, D3 and D4, but there were statistical differences between group D2, D3, D4 and group D1 (F = 44.56, P = 0.00); for ADV, there were no statistical differences among group D2, D3 and D4, but there were statistical differences between group D2, D3, D4 and group D1 (F = 77.52, P = 0.00). For pH ( E ) for RSV, there was no statistical differences between group E5 and E6, and so was between group E3 and E4, but there were statistical differences among group (E5, E6), group (E3, E4), group E1 and group E2 (F = 85.36, P = 0.00); for ADV, there was no statistical differences between group E5 and E6, and so was between group E3 and E4, but there were statistical differences among group (E5, E6), group (E3, E4), group E1 and group E2 (F = 1370.60, P = 0.00).

    Techniques Used: Quantitative RT-PCR

    Related Articles

    Plasmid Preparation:

    Article Title: Lack of nuclear translocation of cytoplasmic domains of IL-2/IL-15 receptor subunits
    Article Snippet: The cDNA encoding the LGV protein was digested from pLGV-N, which is a modified version of pLGV [ ], with Xmn I and Bam H I and inserted into Sma I- and Bam H I-cleaved pCR2.1 plasmid containing the C-terminal region of mouse IL-2Rβ cDNA. .. The resultant plasmid was digested with Bam H I, the ends were filled-in with T4 DNA polymerase (DNA Blunting kit, Takara), and subsequently digested with Apa I. .. The cDNA encoding the fusion protein of the C-terminal region of mouse IL-2Rβ and LGV was ligated into the pMX vector [ ] together with the cDNA encoding the amino-terminal region of mouse IL-2Rβ derived from mIL-2Rβ WT/pBS.

    Article Title: A Novel Recombinant Peste des Petits Ruminants-Canine Adenovirus Vaccine Elicits Long-Lasting Neutralizing Antibody Response against PPR in Goats
    Article Snippet: The H sequence was released with KpnI and XhoI from pMD18-T-H and cloned into the KpnI/XhoI sites of pVAX1 to generate pVAX-H. .. The MluI /HaeII fragment of pVAX-H, containing the H cDNA expression cassette driven by the cytomegalovirus (CMV) immediate early promoter with BGH polyadenylation signals at the other end of the gene, was blunted using a DNA blunting kit (TaKaRa) and directionally cloned into the SspI site of pVAX-ΔE3, forming the pVAXΔE3-H vector. .. The NruI and SalI double digest pVAXΔE3-H fragment containing the H expression cassette flanked with residual E3 sequences was cloned back into pPolyII-CAV-2 by replacing the fragment between the NruI and SalI sites, forming the final pPolyII-CAV-ΔE3-H plasmid ( ).

    Article Title: NF-YC Functions as a Corepressor of Agonist-bound Mineralocorticoid Receptor *
    Article Snippet: In detail, MR fragment was first obtained by PCR amplification with primers containing oligonucleotide linkers of restriction enzyme sites (XmaI-SalI), followed by TA cloning into pCRII-TOPO vector (Invitrogen). .. The pCRII-TOPO-MR-(602–984) construct was then digested with XmaI-SalI and subcloned into pGBKT7 yeast expression vector. pGADT7-NF-YC was cloned as an MR interacting protein by using a yeast two-hybrid system. pcDNA3.1/His-NF-YC was constructed by inserting the 2.1-kb EcoRI fragment of pGADT7-NF-YC from the 3.5-kb original cDNA into the EcoRI site of pcDNA3.1/His vector (Invitrogen). pcDNA3.1/His-NF-YC-(1–115) was constructed by using DNA Blunting Kit (TaKaRa) after excluding the small ClaI-XhoI fragment of pcDNA3.1/His-NF-YC. pcDNA3.1/His-NF-YC-(116–335) was constructed by using the DNA Blunting Kit (TaKaRa) after excluding the small BamHI-ClaI fragment of pcDNA3.1/His-NF-YC. .. Gal4-DBD-NF-YC mammalian expression plasmid of Gal4-DBD-NF-YC (pM-NF-YC) was constructed by inserting the EcoRI fragment of pcDNA3.1/His-NF-YC into the EcoRI site of pM vector. pEGFP-NF-YC was constructed by inserting the EcoRI fragment of pGADT7-NF-YC into the EcoRI site of pEGFP vector.

    Article Title: LINE-1 hypomethylation status of circulating cell-free DNA in plasma as a biomarker for colorectal cancer
    Article Snippet: Methylated control plasmid was digested by Hind III and blunted with DNA Blunting Kit (TAKARA BIO), and then digested by Xho I. .. After the BamH I site of unmethylated control plasmid being destroyed by blunting with DNA Blunting Kit and self-ligation, unmethylated control plasmid was digested by EcoR V and Xho I. .. The Xho I and blunted Hind III fragment of methylated control DNA was ligated into the Xho I and EcoR V site of unmethylated control plasmid vector.

    Polymerase Chain Reaction:

    Article Title: Regulatory RNAs and the HptB/RetS signalling pathways fine-tune Pseudomonas aeruginosa pathogenesis
    Article Snippet: The hptB gene was amplified by PCR from PAK genomic DNA using the primers Bup and Bdown. .. The PCR product was modified using a DNA blunting kit (Takara) and cloned into pUCP18, yielding pUCPhptB . .. The PA3346 and PA3347 genes were amplified by PCR from PAK genomic DNA using the primers.

    Article Title: Chicken CCDC152 shares an NFYB-regulated bidirectional promoter with a growth hormone receptor antisense transcript and inhibits cells proliferation and migration
    Article Snippet: Then 48 h after transfection, cells were lysed with passive lysis buffer, and luciferase activity was measured using the Dual-Luciferase Assay System (Promega) according to the manufacturer's protocols. .. Site mutation constructs and transient transfection assays Various progressive deletion mutants (M1-M8, M1′-M8′) were further constructed using either a DNA blunting kit (Takara, Japan) with restriction enzyme digestion and ligation, or the Site-Directed Mutagenesis Kit (TOYOBO, Japan) with reverse PCR amplification and ligation, according to the manufacturers’ instructions. .. Constructs with point mutation(s) at putative transcription factor binding sites were generated using Ω-PCR [ ].

    Modification:

    Article Title: Regulatory RNAs and the HptB/RetS signalling pathways fine-tune Pseudomonas aeruginosa pathogenesis
    Article Snippet: The hptB gene was amplified by PCR from PAK genomic DNA using the primers Bup and Bdown. .. The PCR product was modified using a DNA blunting kit (Takara) and cloned into pUCP18, yielding pUCPhptB . .. The PA3346 and PA3347 genes were amplified by PCR from PAK genomic DNA using the primers.

    Clone Assay:

    Article Title: Regulatory RNAs and the HptB/RetS signalling pathways fine-tune Pseudomonas aeruginosa pathogenesis
    Article Snippet: The hptB gene was amplified by PCR from PAK genomic DNA using the primers Bup and Bdown. .. The PCR product was modified using a DNA blunting kit (Takara) and cloned into pUCP18, yielding pUCPhptB . .. The PA3346 and PA3347 genes were amplified by PCR from PAK genomic DNA using the primers.

    Article Title: A Novel Recombinant Peste des Petits Ruminants-Canine Adenovirus Vaccine Elicits Long-Lasting Neutralizing Antibody Response against PPR in Goats
    Article Snippet: The H sequence was released with KpnI and XhoI from pMD18-T-H and cloned into the KpnI/XhoI sites of pVAX1 to generate pVAX-H. .. The MluI /HaeII fragment of pVAX-H, containing the H cDNA expression cassette driven by the cytomegalovirus (CMV) immediate early promoter with BGH polyadenylation signals at the other end of the gene, was blunted using a DNA blunting kit (TaKaRa) and directionally cloned into the SspI site of pVAX-ΔE3, forming the pVAXΔE3-H vector. .. The NruI and SalI double digest pVAXΔE3-H fragment containing the H expression cassette flanked with residual E3 sequences was cloned back into pPolyII-CAV-2 by replacing the fragment between the NruI and SalI sites, forming the final pPolyII-CAV-ΔE3-H plasmid ( ).

    Article Title: NF-YC Functions as a Corepressor of Agonist-bound Mineralocorticoid Receptor *
    Article Snippet: In detail, MR fragment was first obtained by PCR amplification with primers containing oligonucleotide linkers of restriction enzyme sites (XmaI-SalI), followed by TA cloning into pCRII-TOPO vector (Invitrogen). .. The pCRII-TOPO-MR-(602–984) construct was then digested with XmaI-SalI and subcloned into pGBKT7 yeast expression vector. pGADT7-NF-YC was cloned as an MR interacting protein by using a yeast two-hybrid system. pcDNA3.1/His-NF-YC was constructed by inserting the 2.1-kb EcoRI fragment of pGADT7-NF-YC from the 3.5-kb original cDNA into the EcoRI site of pcDNA3.1/His vector (Invitrogen). pcDNA3.1/His-NF-YC-(1–115) was constructed by using DNA Blunting Kit (TaKaRa) after excluding the small ClaI-XhoI fragment of pcDNA3.1/His-NF-YC. pcDNA3.1/His-NF-YC-(116–335) was constructed by using the DNA Blunting Kit (TaKaRa) after excluding the small BamHI-ClaI fragment of pcDNA3.1/His-NF-YC. .. Gal4-DBD-NF-YC mammalian expression plasmid of Gal4-DBD-NF-YC (pM-NF-YC) was constructed by inserting the EcoRI fragment of pcDNA3.1/His-NF-YC into the EcoRI site of pM vector. pEGFP-NF-YC was constructed by inserting the EcoRI fragment of pGADT7-NF-YC into the EcoRI site of pEGFP vector.

    Expressing:

    Article Title: A Novel Recombinant Peste des Petits Ruminants-Canine Adenovirus Vaccine Elicits Long-Lasting Neutralizing Antibody Response against PPR in Goats
    Article Snippet: The H sequence was released with KpnI and XhoI from pMD18-T-H and cloned into the KpnI/XhoI sites of pVAX1 to generate pVAX-H. .. The MluI /HaeII fragment of pVAX-H, containing the H cDNA expression cassette driven by the cytomegalovirus (CMV) immediate early promoter with BGH polyadenylation signals at the other end of the gene, was blunted using a DNA blunting kit (TaKaRa) and directionally cloned into the SspI site of pVAX-ΔE3, forming the pVAXΔE3-H vector. .. The NruI and SalI double digest pVAXΔE3-H fragment containing the H expression cassette flanked with residual E3 sequences was cloned back into pPolyII-CAV-2 by replacing the fragment between the NruI and SalI sites, forming the final pPolyII-CAV-ΔE3-H plasmid ( ).

    Article Title: NF-YC Functions as a Corepressor of Agonist-bound Mineralocorticoid Receptor *
    Article Snippet: In detail, MR fragment was first obtained by PCR amplification with primers containing oligonucleotide linkers of restriction enzyme sites (XmaI-SalI), followed by TA cloning into pCRII-TOPO vector (Invitrogen). .. The pCRII-TOPO-MR-(602–984) construct was then digested with XmaI-SalI and subcloned into pGBKT7 yeast expression vector. pGADT7-NF-YC was cloned as an MR interacting protein by using a yeast two-hybrid system. pcDNA3.1/His-NF-YC was constructed by inserting the 2.1-kb EcoRI fragment of pGADT7-NF-YC from the 3.5-kb original cDNA into the EcoRI site of pcDNA3.1/His vector (Invitrogen). pcDNA3.1/His-NF-YC-(1–115) was constructed by using DNA Blunting Kit (TaKaRa) after excluding the small ClaI-XhoI fragment of pcDNA3.1/His-NF-YC. pcDNA3.1/His-NF-YC-(116–335) was constructed by using the DNA Blunting Kit (TaKaRa) after excluding the small BamHI-ClaI fragment of pcDNA3.1/His-NF-YC. .. Gal4-DBD-NF-YC mammalian expression plasmid of Gal4-DBD-NF-YC (pM-NF-YC) was constructed by inserting the EcoRI fragment of pcDNA3.1/His-NF-YC into the EcoRI site of pM vector. pEGFP-NF-YC was constructed by inserting the EcoRI fragment of pGADT7-NF-YC into the EcoRI site of pEGFP vector.

    Mutagenesis:

    Article Title: Chicken CCDC152 shares an NFYB-regulated bidirectional promoter with a growth hormone receptor antisense transcript and inhibits cells proliferation and migration
    Article Snippet: Then 48 h after transfection, cells were lysed with passive lysis buffer, and luciferase activity was measured using the Dual-Luciferase Assay System (Promega) according to the manufacturer's protocols. .. Site mutation constructs and transient transfection assays Various progressive deletion mutants (M1-M8, M1′-M8′) were further constructed using either a DNA blunting kit (Takara, Japan) with restriction enzyme digestion and ligation, or the Site-Directed Mutagenesis Kit (TOYOBO, Japan) with reverse PCR amplification and ligation, according to the manufacturers’ instructions. .. Constructs with point mutation(s) at putative transcription factor binding sites were generated using Ω-PCR [ ].

    Construct:

    Article Title: Chicken CCDC152 shares an NFYB-regulated bidirectional promoter with a growth hormone receptor antisense transcript and inhibits cells proliferation and migration
    Article Snippet: Then 48 h after transfection, cells were lysed with passive lysis buffer, and luciferase activity was measured using the Dual-Luciferase Assay System (Promega) according to the manufacturer's protocols. .. Site mutation constructs and transient transfection assays Various progressive deletion mutants (M1-M8, M1′-M8′) were further constructed using either a DNA blunting kit (Takara, Japan) with restriction enzyme digestion and ligation, or the Site-Directed Mutagenesis Kit (TOYOBO, Japan) with reverse PCR amplification and ligation, according to the manufacturers’ instructions. .. Constructs with point mutation(s) at putative transcription factor binding sites were generated using Ω-PCR [ ].

    Article Title: NF-YC Functions as a Corepressor of Agonist-bound Mineralocorticoid Receptor *
    Article Snippet: In detail, MR fragment was first obtained by PCR amplification with primers containing oligonucleotide linkers of restriction enzyme sites (XmaI-SalI), followed by TA cloning into pCRII-TOPO vector (Invitrogen). .. The pCRII-TOPO-MR-(602–984) construct was then digested with XmaI-SalI and subcloned into pGBKT7 yeast expression vector. pGADT7-NF-YC was cloned as an MR interacting protein by using a yeast two-hybrid system. pcDNA3.1/His-NF-YC was constructed by inserting the 2.1-kb EcoRI fragment of pGADT7-NF-YC from the 3.5-kb original cDNA into the EcoRI site of pcDNA3.1/His vector (Invitrogen). pcDNA3.1/His-NF-YC-(1–115) was constructed by using DNA Blunting Kit (TaKaRa) after excluding the small ClaI-XhoI fragment of pcDNA3.1/His-NF-YC. pcDNA3.1/His-NF-YC-(116–335) was constructed by using the DNA Blunting Kit (TaKaRa) after excluding the small BamHI-ClaI fragment of pcDNA3.1/His-NF-YC. .. Gal4-DBD-NF-YC mammalian expression plasmid of Gal4-DBD-NF-YC (pM-NF-YC) was constructed by inserting the EcoRI fragment of pcDNA3.1/His-NF-YC into the EcoRI site of pM vector. pEGFP-NF-YC was constructed by inserting the EcoRI fragment of pGADT7-NF-YC into the EcoRI site of pEGFP vector.

    Transfection:

    Article Title: Chicken CCDC152 shares an NFYB-regulated bidirectional promoter with a growth hormone receptor antisense transcript and inhibits cells proliferation and migration
    Article Snippet: Then 48 h after transfection, cells were lysed with passive lysis buffer, and luciferase activity was measured using the Dual-Luciferase Assay System (Promega) according to the manufacturer's protocols. .. Site mutation constructs and transient transfection assays Various progressive deletion mutants (M1-M8, M1′-M8′) were further constructed using either a DNA blunting kit (Takara, Japan) with restriction enzyme digestion and ligation, or the Site-Directed Mutagenesis Kit (TOYOBO, Japan) with reverse PCR amplification and ligation, according to the manufacturers’ instructions. .. Constructs with point mutation(s) at putative transcription factor binding sites were generated using Ω-PCR [ ].

    Ligation:

    Article Title: Chicken CCDC152 shares an NFYB-regulated bidirectional promoter with a growth hormone receptor antisense transcript and inhibits cells proliferation and migration
    Article Snippet: Then 48 h after transfection, cells were lysed with passive lysis buffer, and luciferase activity was measured using the Dual-Luciferase Assay System (Promega) according to the manufacturer's protocols. .. Site mutation constructs and transient transfection assays Various progressive deletion mutants (M1-M8, M1′-M8′) were further constructed using either a DNA blunting kit (Takara, Japan) with restriction enzyme digestion and ligation, or the Site-Directed Mutagenesis Kit (TOYOBO, Japan) with reverse PCR amplification and ligation, according to the manufacturers’ instructions. .. Constructs with point mutation(s) at putative transcription factor binding sites were generated using Ω-PCR [ ].

    Amplification:

    Article Title: Chicken CCDC152 shares an NFYB-regulated bidirectional promoter with a growth hormone receptor antisense transcript and inhibits cells proliferation and migration
    Article Snippet: Then 48 h after transfection, cells were lysed with passive lysis buffer, and luciferase activity was measured using the Dual-Luciferase Assay System (Promega) according to the manufacturer's protocols. .. Site mutation constructs and transient transfection assays Various progressive deletion mutants (M1-M8, M1′-M8′) were further constructed using either a DNA blunting kit (Takara, Japan) with restriction enzyme digestion and ligation, or the Site-Directed Mutagenesis Kit (TOYOBO, Japan) with reverse PCR amplification and ligation, according to the manufacturers’ instructions. .. Constructs with point mutation(s) at putative transcription factor binding sites were generated using Ω-PCR [ ].

    Immunoprecipitation:

    Article Title: Bean Metal-Responsive Element-Binding Transcription Factor Confers Cadmium Resistance in Tobacco 1
    Article Snippet: Four-week-old 35S : PvMTF-1-GFP transgenic tobacco seedlings were subjected to ChIP assay using anti-GFP antibody (ab290, Abcam) according to the instruction of the EpiQuik Plant ChIP Kit (Epigentek). .. Immunoprecipitated DNA products were blunted using DNA Blunting Kit (TaKaRa), and then one A tail was added to the 3′ end of blunt DNA according to the instructions for the DNA A-Tailing Kit (TaKaRa). .. The resulting DNA products were cloned into pMD18T-vector (TaKaRa) and sequenced.

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    • plant genomic dna extraction kit  (TaKaRa)
    96
    TaKaRa plant genomic dna extraction kit
    Generation and molecular analysis of transgenic plants. (A) Schematic representation of the pANDA35HK- dsHaAK expression cassettes used for <t>Arabidopsis</t> transformation. 35S pro, CaMV 35S promoter; HPT, hygromycin phosphotralsferase gene; NPT II, neomycin phosphotransferase II gene; HaAK , cDNA sequence of AK gene from H. armigera ; RB, right border; LB, left border; (B) Detection of HaAK in non-transformed control and transgenic plants by PCR. A 1,068 bp fragment of HaAK was amplified and 18s rDNA was served as a control. Lane M, <t>DNA</t> marker DL2,000; Lane 1, untransformed control; Lane 2-8, the transformants; (C) The wild and transgenic plants were grown on kanamycin-containing medium. The homozygous single-copy transgenic plants were selected through Mendelian segregation; (D) Northern blot detection of HaAK dsRNA in control and different transgenic lines. Lane 1, the non-transformed control; Lane 2-9, the transformants (line AtdsHaAK-2 , AtdsHaAK-3, AtdsHaAK-6, AtdsHaAK-7, AtdsHaAK-8, AtdsHaAK-9, AtdsHaAK-11 and AtdsHaAK-12 ); Actin1 was used as a reference and the DIG labeled probe for Actin1 was obtained by PCR using primers as described for RT-PCR. (E) RT-PCR amplification of HaAK in control and transgenic plants. A 1,068 bp fragment of HaAK was amplified and Arabidopsis Actin1 gene was used as a control. Lane 1, untransformed plant; Lane 2-9, the transformants (line AtdsHaAK-2 , AtdsHaAK-3, AtdsHaAK-6, AtdsHaAK-7, AtdsHaAK-8, AtdsHaAK-9, AtdsHaAK-11, AtdsHaAK-12 ); (F) Northern blot detection of small RNA fragments of HaAK in transgenic plants leaves . Lane 1, non-transformed plant; Lane 2-7, the transformants.
    Plant Genomic Dna Extraction Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plant genomic dna extraction kit/product/TaKaRa
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    plant genomic dna extraction kit - by Bioz Stars, 2021-03
    96/100 stars
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    dna extraction kit  (TaKaRa)
    99
    TaKaRa dna extraction kit
    IRF1 inhibits PGC1α expression via directly binding to its promoter region. a After being co-transfected with pRL-TK vector and pGL3-basic or recombinant reporter plasmids containing various fragments of PGC1α promoter region, <t>H9c2</t> cells were treated with control or HP and then harvested for dual-luciferase reporter assay. The firefly luciferase activity was normalized against Renilla activity. b H9c2 cells were co-transfected with pRL-TK vector and pGL3-PGC1α-P3 or pGL3-PGC1α-M3 (containing the mutant bases of pGL3-PGC1α-P3 underlined in the sequencing results), treated with control or HP and harvested for luciferase assay. c , d H9c2 cells were treated with control or HP for 24 h and harvested for ChIP assay. IRF1 antibody was immunoprecipitated with chromatin <t>DNA</t> fragments, taking IgG as a negative control. The precipitated DNA was amplified by PCR c and qPCR d using primers that cover the PGC1α promoter region (−709 to −511), taking primers that cover the region (−1711 to −1598) as a negative control. e , f qPCR and representative western blot analysis of IRF1 and PGC1α expressions in cells treated with siPit1, siPit2, or both in the presence or absence of HP. g , h H9c2 cells were treated with control or HP, and H3K9 acetylation was assayed using ChIP. i qPCR analysis of histone acetyltransferase genes expression in H9c2 cells treated with control or HP. j – l qPCR and western blot analysis of IRF1 and PGC1α expressions in cells treated with a P300/CBP inhibitor (C646, 10 μM) or a pan-histone acetyltransferase inhibitor (anacardic acid, AA,10 μM) in the presence or absence of HP. Data are shown as mean ± SD and were analyzed by a two-tailed unpaired t -test ( a – d , h , i ) or one-way ANOVA ( e , f , j , k ). n = 3 biologically independent experiments ( a , b , d – f , h – k ). * P
    Dna Extraction Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    cdna synthesis  (TaKaRa)
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    Increased TRP1 and TRP2 expressions in SRA-deficient B16 cells. B16-shCtrl and B16-SRAi cells were processed for <t>RNA</t> extraction, <t>cDNA</t> synthesis, and real-time PCR for the measurement of MITF, TYR, TRP1, TRP2, endothelin-1, and MC1R (n = 3 and 3, respectively; error bars represent standard errors; * indicates p
    Cdna Synthesis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Generation and molecular analysis of transgenic plants. (A) Schematic representation of the pANDA35HK- dsHaAK expression cassettes used for Arabidopsis transformation. 35S pro, CaMV 35S promoter; HPT, hygromycin phosphotralsferase gene; NPT II, neomycin phosphotransferase II gene; HaAK , cDNA sequence of AK gene from H. armigera ; RB, right border; LB, left border; (B) Detection of HaAK in non-transformed control and transgenic plants by PCR. A 1,068 bp fragment of HaAK was amplified and 18s rDNA was served as a control. Lane M, DNA marker DL2,000; Lane 1, untransformed control; Lane 2-8, the transformants; (C) The wild and transgenic plants were grown on kanamycin-containing medium. The homozygous single-copy transgenic plants were selected through Mendelian segregation; (D) Northern blot detection of HaAK dsRNA in control and different transgenic lines. Lane 1, the non-transformed control; Lane 2-9, the transformants (line AtdsHaAK-2 , AtdsHaAK-3, AtdsHaAK-6, AtdsHaAK-7, AtdsHaAK-8, AtdsHaAK-9, AtdsHaAK-11 and AtdsHaAK-12 ); Actin1 was used as a reference and the DIG labeled probe for Actin1 was obtained by PCR using primers as described for RT-PCR. (E) RT-PCR amplification of HaAK in control and transgenic plants. A 1,068 bp fragment of HaAK was amplified and Arabidopsis Actin1 gene was used as a control. Lane 1, untransformed plant; Lane 2-9, the transformants (line AtdsHaAK-2 , AtdsHaAK-3, AtdsHaAK-6, AtdsHaAK-7, AtdsHaAK-8, AtdsHaAK-9, AtdsHaAK-11, AtdsHaAK-12 ); (F) Northern blot detection of small RNA fragments of HaAK in transgenic plants leaves . Lane 1, non-transformed plant; Lane 2-7, the transformants.

    Journal: International Journal of Biological Sciences

    Article Title: Silencing the HaAK Gene by Transgenic Plant-Mediated RNAi Impairs Larval Growth of Helicoverpa armigera

    doi: 10.7150/ijbs.10468

    Figure Lengend Snippet: Generation and molecular analysis of transgenic plants. (A) Schematic representation of the pANDA35HK- dsHaAK expression cassettes used for Arabidopsis transformation. 35S pro, CaMV 35S promoter; HPT, hygromycin phosphotralsferase gene; NPT II, neomycin phosphotransferase II gene; HaAK , cDNA sequence of AK gene from H. armigera ; RB, right border; LB, left border; (B) Detection of HaAK in non-transformed control and transgenic plants by PCR. A 1,068 bp fragment of HaAK was amplified and 18s rDNA was served as a control. Lane M, DNA marker DL2,000; Lane 1, untransformed control; Lane 2-8, the transformants; (C) The wild and transgenic plants were grown on kanamycin-containing medium. The homozygous single-copy transgenic plants were selected through Mendelian segregation; (D) Northern blot detection of HaAK dsRNA in control and different transgenic lines. Lane 1, the non-transformed control; Lane 2-9, the transformants (line AtdsHaAK-2 , AtdsHaAK-3, AtdsHaAK-6, AtdsHaAK-7, AtdsHaAK-8, AtdsHaAK-9, AtdsHaAK-11 and AtdsHaAK-12 ); Actin1 was used as a reference and the DIG labeled probe for Actin1 was obtained by PCR using primers as described for RT-PCR. (E) RT-PCR amplification of HaAK in control and transgenic plants. A 1,068 bp fragment of HaAK was amplified and Arabidopsis Actin1 gene was used as a control. Lane 1, untransformed plant; Lane 2-9, the transformants (line AtdsHaAK-2 , AtdsHaAK-3, AtdsHaAK-6, AtdsHaAK-7, AtdsHaAK-8, AtdsHaAK-9, AtdsHaAK-11, AtdsHaAK-12 ); (F) Northern blot detection of small RNA fragments of HaAK in transgenic plants leaves . Lane 1, non-transformed plant; Lane 2-7, the transformants.

    Article Snippet: Molecular analysis of transgenic plants Total genomic DNA was isolated from the leaf tissues of Arabidopsis using a plant genomic DNA extraction kit (Takara, China).

    Techniques: Transgenic Assay, Expressing, Transformation Assay, Sequencing, Polymerase Chain Reaction, Amplification, Marker, Northern Blot, Labeling, Reverse Transcription Polymerase Chain Reaction

    IRF1 inhibits PGC1α expression via directly binding to its promoter region. a After being co-transfected with pRL-TK vector and pGL3-basic or recombinant reporter plasmids containing various fragments of PGC1α promoter region, H9c2 cells were treated with control or HP and then harvested for dual-luciferase reporter assay. The firefly luciferase activity was normalized against Renilla activity. b H9c2 cells were co-transfected with pRL-TK vector and pGL3-PGC1α-P3 or pGL3-PGC1α-M3 (containing the mutant bases of pGL3-PGC1α-P3 underlined in the sequencing results), treated with control or HP and harvested for luciferase assay. c , d H9c2 cells were treated with control or HP for 24 h and harvested for ChIP assay. IRF1 antibody was immunoprecipitated with chromatin DNA fragments, taking IgG as a negative control. The precipitated DNA was amplified by PCR c and qPCR d using primers that cover the PGC1α promoter region (−709 to −511), taking primers that cover the region (−1711 to −1598) as a negative control. e , f qPCR and representative western blot analysis of IRF1 and PGC1α expressions in cells treated with siPit1, siPit2, or both in the presence or absence of HP. g , h H9c2 cells were treated with control or HP, and H3K9 acetylation was assayed using ChIP. i qPCR analysis of histone acetyltransferase genes expression in H9c2 cells treated with control or HP. j – l qPCR and western blot analysis of IRF1 and PGC1α expressions in cells treated with a P300/CBP inhibitor (C646, 10 μM) or a pan-histone acetyltransferase inhibitor (anacardic acid, AA,10 μM) in the presence or absence of HP. Data are shown as mean ± SD and were analyzed by a two-tailed unpaired t -test ( a – d , h , i ) or one-way ANOVA ( e , f , j , k ). n = 3 biologically independent experiments ( a , b , d – f , h – k ). * P

    Journal: Nature Communications

    Article Title: IRF1-mediated downregulation of PGC1α contributes to cardiorenal syndrome type 4

    doi: 10.1038/s41467-020-18519-0

    Figure Lengend Snippet: IRF1 inhibits PGC1α expression via directly binding to its promoter region. a After being co-transfected with pRL-TK vector and pGL3-basic or recombinant reporter plasmids containing various fragments of PGC1α promoter region, H9c2 cells were treated with control or HP and then harvested for dual-luciferase reporter assay. The firefly luciferase activity was normalized against Renilla activity. b H9c2 cells were co-transfected with pRL-TK vector and pGL3-PGC1α-P3 or pGL3-PGC1α-M3 (containing the mutant bases of pGL3-PGC1α-P3 underlined in the sequencing results), treated with control or HP and harvested for luciferase assay. c , d H9c2 cells were treated with control or HP for 24 h and harvested for ChIP assay. IRF1 antibody was immunoprecipitated with chromatin DNA fragments, taking IgG as a negative control. The precipitated DNA was amplified by PCR c and qPCR d using primers that cover the PGC1α promoter region (−709 to −511), taking primers that cover the region (−1711 to −1598) as a negative control. e , f qPCR and representative western blot analysis of IRF1 and PGC1α expressions in cells treated with siPit1, siPit2, or both in the presence or absence of HP. g , h H9c2 cells were treated with control or HP, and H3K9 acetylation was assayed using ChIP. i qPCR analysis of histone acetyltransferase genes expression in H9c2 cells treated with control or HP. j – l qPCR and western blot analysis of IRF1 and PGC1α expressions in cells treated with a P300/CBP inhibitor (C646, 10 μM) or a pan-histone acetyltransferase inhibitor (anacardic acid, AA,10 μM) in the presence or absence of HP. Data are shown as mean ± SD and were analyzed by a two-tailed unpaired t -test ( a – d , h , i ) or one-way ANOVA ( e , f , j , k ). n = 3 biologically independent experiments ( a , b , d – f , h – k ). * P

    Article Snippet: Mitochondrial DNA analysisTotal DNA, including chromosomal (B2M) and mitochondrial (D-loop) DNA, was extracted from H9c2 cells using a DNA extraction kit (Takara) following the manufacturer’s instructions.

    Techniques: Expressing, Binding Assay, Transfection, Plasmid Preparation, Recombinant, Luciferase, Reporter Assay, Activity Assay, Mutagenesis, Sequencing, Chromatin Immunoprecipitation, Immunoprecipitation, Negative Control, Amplification, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Western Blot, Two Tailed Test

    HP induces PGC1α reduction and mitochondrial energy metabolism dysfunction in vitro. a H9c2 cells were treated with urea (1.2 mg/ml), serum creatinine (Scr, 80 μg/ml), uric acid (UA, 80 μg/ml), parathyroid hormone (PTH, 90 ng/ml), trimethylamine- N -oxide (TMAO, 0.751 μg/ml), HP (84 μg/ml), FGF23 (20 ng/ml), β2-microglobulin (B2M, 20 μg/ml), indoxyl sulfate (IS, 125 μg/ml), p -cresyl sulfate (PCS, 47 μg/ml), and indole-3-acetic acid (IAA, 3.5 μg/ml), separately. mRNA was extracted for qPCR analysis of PGC1α expression. b , c qPCR and western blot analysis of PGC1α expression in H9c2 cells incubated with the serum of healthy donors or CKD patients for 24 h. d – g qPCR and representative western blot analysis of PGC1α expression in H9c2 cells treated with control, various doses of HP for 24 h or HP (8.4 mg/dl) for various durations. h – p Oxygen consumption rate (OCR) ( h ), relative ATP level ( i ), relative mRNA expression of FAO genes ( j ), relative mRNA expression of OXPHO genes ( k ), relative mRNA expression of glycolysis genes ( l ), relative mitochondrial DNA copy number ( m ), ROS production ( n ), and monomer and aggregate JC-1 ( o flow cytometry; p representative images of laser scanning confocal microscopy. Scale bar, 10 μm) in H9c2 cells treated with control or HP for 24 h. Data are shown as mean ± SD and were analyzed by a two-tailed unpaired t -test ( a – c , h – o ) or one-way ANOVA ( d – g ). n = 3 ( a – g , i – p ) or n = 4 ( h ) biologically independent experiments. * P

    Journal: Nature Communications

    Article Title: IRF1-mediated downregulation of PGC1α contributes to cardiorenal syndrome type 4

    doi: 10.1038/s41467-020-18519-0

    Figure Lengend Snippet: HP induces PGC1α reduction and mitochondrial energy metabolism dysfunction in vitro. a H9c2 cells were treated with urea (1.2 mg/ml), serum creatinine (Scr, 80 μg/ml), uric acid (UA, 80 μg/ml), parathyroid hormone (PTH, 90 ng/ml), trimethylamine- N -oxide (TMAO, 0.751 μg/ml), HP (84 μg/ml), FGF23 (20 ng/ml), β2-microglobulin (B2M, 20 μg/ml), indoxyl sulfate (IS, 125 μg/ml), p -cresyl sulfate (PCS, 47 μg/ml), and indole-3-acetic acid (IAA, 3.5 μg/ml), separately. mRNA was extracted for qPCR analysis of PGC1α expression. b , c qPCR and western blot analysis of PGC1α expression in H9c2 cells incubated with the serum of healthy donors or CKD patients for 24 h. d – g qPCR and representative western blot analysis of PGC1α expression in H9c2 cells treated with control, various doses of HP for 24 h or HP (8.4 mg/dl) for various durations. h – p Oxygen consumption rate (OCR) ( h ), relative ATP level ( i ), relative mRNA expression of FAO genes ( j ), relative mRNA expression of OXPHO genes ( k ), relative mRNA expression of glycolysis genes ( l ), relative mitochondrial DNA copy number ( m ), ROS production ( n ), and monomer and aggregate JC-1 ( o flow cytometry; p representative images of laser scanning confocal microscopy. Scale bar, 10 μm) in H9c2 cells treated with control or HP for 24 h. Data are shown as mean ± SD and were analyzed by a two-tailed unpaired t -test ( a – c , h – o ) or one-way ANOVA ( d – g ). n = 3 ( a – g , i – p ) or n = 4 ( h ) biologically independent experiments. * P

    Article Snippet: Mitochondrial DNA analysisTotal DNA, including chromosomal (B2M) and mitochondrial (D-loop) DNA, was extracted from H9c2 cells using a DNA extraction kit (Takara) following the manufacturer’s instructions.

    Techniques: In Vitro, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Incubation, Flow Cytometry, Confocal Microscopy, Two Tailed Test

    PI3K specific inhibitor LY294002 inhibits MDV replication. CEFs were pre-incubated with LY294002 (20 μM), LY303511 (30 μM) or wortmannin (500 μM) for 1 h, infected with MDV-LZ1309 at 0.1 MOI and analyzed for virus replication at 24, 48, 72, and 96 hpi. (A) MDV plaque quantification. (B) TaqMan real-time PCR detection of viral DNA. The data represent the mean ± SD of three independent experiments. One-way ANOVA, ∗ P

    Journal: Frontiers in Microbiology

    Article Title: Marek’s Disease Virus Activates the PI3K/Akt Pathway Through Interaction of Its Protein Meq With the P85 Subunit of PI3K to Promote Viral Replication

    doi: 10.3389/fmicb.2018.02547

    Figure Lengend Snippet: PI3K specific inhibitor LY294002 inhibits MDV replication. CEFs were pre-incubated with LY294002 (20 μM), LY303511 (30 μM) or wortmannin (500 μM) for 1 h, infected with MDV-LZ1309 at 0.1 MOI and analyzed for virus replication at 24, 48, 72, and 96 hpi. (A) MDV plaque quantification. (B) TaqMan real-time PCR detection of viral DNA. The data represent the mean ± SD of three independent experiments. One-way ANOVA, ∗ P

    Article Snippet: The following primers were used: PIK3R1 forward: 5′-CCTCGAGGATGATGGCTGAAGGATATCA-3′ and reverse: 5′-CGGATCCGTCAATCGCCTCTGCTGTGCAT-3′; PIK3R2 forward: 5′-CCTCGAGGATGGGCAGAACTGACGGATTC-3′ and reverse: 5′-CGGATCCGTCATATGGATGGTGGCTGGGA-3′; And the meq gene of the MDV LZ1309 strain (GenBank accession number KX966280.1 ) were amplified by PCR which DNA was extracted using DNA Extraction Kit (Takara, Dalian, China) from LZ1309-infected cells.

    Techniques: Incubation, Infection, Real-time Polymerase Chain Reaction

    Increased TRP1 and TRP2 expressions in SRA-deficient B16 cells. B16-shCtrl and B16-SRAi cells were processed for RNA extraction, cDNA synthesis, and real-time PCR for the measurement of MITF, TYR, TRP1, TRP2, endothelin-1, and MC1R (n = 3 and 3, respectively; error bars represent standard errors; * indicates p

    Journal: PLoS ONE

    Article Title: Targeting steroid receptor RNA activator (SRA), a long non-coding RNA, enhances melanogenesis through activation of TRP1 and inhibition of p38 phosphorylation

    doi: 10.1371/journal.pone.0237577

    Figure Lengend Snippet: Increased TRP1 and TRP2 expressions in SRA-deficient B16 cells. B16-shCtrl and B16-SRAi cells were processed for RNA extraction, cDNA synthesis, and real-time PCR for the measurement of MITF, TYR, TRP1, TRP2, endothelin-1, and MC1R (n = 3 and 3, respectively; error bars represent standard errors; * indicates p

    Article Snippet: One microgram of total RNA was used for cDNA synthesis by using PrimeScript RT Reagent Kit (RR037A, TaKaRa).

    Techniques: RNA Extraction, Real-time Polymerase Chain Reaction