px330 vector  (New England Biolabs)


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    New England Biolabs px330 vector
    Generation and validation of WNK1 knockout (KO) cell lines. A : mismatch-specific endonuclease assay. Genomic PCR (gPCR) products spanning exon 1 of WNK1 were amplified from the template of a heterogeneous population of HEK293T cells transfected with <t>px330-WNK1</t>
    Px330 Vector, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/px330 vector/product/New England Biolabs
    Average 91 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    px330 vector - by Bioz Stars, 2020-07
    91/100 stars

    Images

    1) Product Images from "Generation of WNK1 knockout cell lines by CRISPR/Cas-mediated genome editing"

    Article Title: Generation of WNK1 knockout cell lines by CRISPR/Cas-mediated genome editing

    Journal: American Journal of Physiology - Renal Physiology

    doi: 10.1152/ajprenal.00612.2014

    Generation and validation of WNK1 knockout (KO) cell lines. A : mismatch-specific endonuclease assay. Genomic PCR (gPCR) products spanning exon 1 of WNK1 were amplified from the template of a heterogeneous population of HEK293T cells transfected with px330-WNK1
    Figure Legend Snippet: Generation and validation of WNK1 knockout (KO) cell lines. A : mismatch-specific endonuclease assay. Genomic PCR (gPCR) products spanning exon 1 of WNK1 were amplified from the template of a heterogeneous population of HEK293T cells transfected with px330-WNK1

    Techniques Used: Knock-Out, Polymerase Chain Reaction, Amplification, Transfection

    Related Articles

    Clone Assay:

    Article Title: Transcriptional activation of fucosyltransferase (FUT) genes using the CRISPR-dCas9-VPR technology reveals potent N-glycome alterations in colorectal cancer cells
    Article Snippet: .. The PCR product encoding the Puromycin marker was cloned into the pX330 vector after backbone linearization using two neighboring Sma I restriction sites (New England BioLabs) and InFusion cloning (Clontech, California, USA). .. Next, the Cas9 coding sequence was removed from the vector backbone by Nco I and Eco RI (New England BioLabs) digests.

    Article Title: Transcriptional activation of fucosyltransferase (FUT) genes using the CRISPR-dCas9-VPR technology reveals potent N-glycome alterations in colorectal cancer cells
    Article Snippet: .. The PCR product encoding the Puromycin marker was cloned into the pX330 vector after backbone linearization using two neighboring Sma I restriction sites (New England BioLabs) and InFusion cloning (Clontech, California, USA). .. Next, the Cas9 coding sequence was removed from the vector backbone by Nco I and Eco RI (New England BioLabs) digests.

    Transfection:

    Article Title: Drug‐induced increase in lysobisphosphatidic acid reduces the cholesterol overload in Niemann–Pick type C cells and mice
    Article Snippet: .. The sequences were used to insert the target sequence into the pX330 vector using Golden Gate Assembly (New England Biolabs) and transfected into cells. .. Knockout clones were isolated by serial dilution and confirmed by RT–PCR, Western blotting and filipin staining.

    Article Title: Cyclodextrin triggers MCOLN1-dependent endo-lysosome secretion in Niemann-Pick type C cells [S]
    Article Snippet: .. The sequences were used to insert the target sequence into the pX330 vector using Golden Gate Assembly (New England Biolabs) and transfected into cells. .. KO clones were isolated by serial dilution and confirmed by RT-PCR, Western blotting, and filipin staining.

    Amplification:

    Article Title: Genome editing reveals a role for OCT4 in human embryogenesis
    Article Snippet: .. The sgRNA sequence from the correctly targeted px330 vector was amplified using the Q5 hot start high fidelity DNA polymerase (NEB; M0493) and the PCR product was in vitro transcribed using the MEGAshortscript T7 kit (ThermoFisher Scientific; AM1354) and purified using the Zymo RNA Clean & Concentrator columns (Zymo Research; R1017) The sgRNA and Cas9 mRNA (TriLink Biotechnologies; L61256) and recombinant Cas9 protein (Toolgen; TGEN CP1) were individually re-suspended in RNase-free water, aliquoted and stored at −80 °C until use. .. Prior to microinjection, the ribonucleoprotein complex was prepared by centrifuging the Cas9 protein for 1 min at 14,000 r.p.m. at 4 °C and transferring the supernatant to a fresh tube containing the sgRNA.

    In Vitro:

    Article Title: Genome editing reveals a role for OCT4 in human embryogenesis
    Article Snippet: .. The sgRNA sequence from the correctly targeted px330 vector was amplified using the Q5 hot start high fidelity DNA polymerase (NEB; M0493) and the PCR product was in vitro transcribed using the MEGAshortscript T7 kit (ThermoFisher Scientific; AM1354) and purified using the Zymo RNA Clean & Concentrator columns (Zymo Research; R1017) The sgRNA and Cas9 mRNA (TriLink Biotechnologies; L61256) and recombinant Cas9 protein (Toolgen; TGEN CP1) were individually re-suspended in RNase-free water, aliquoted and stored at −80 °C until use. .. Prior to microinjection, the ribonucleoprotein complex was prepared by centrifuging the Cas9 protein for 1 min at 14,000 r.p.m. at 4 °C and transferring the supernatant to a fresh tube containing the sgRNA.

    Polymerase Chain Reaction:

    Article Title: Transcriptional activation of fucosyltransferase (FUT) genes using the CRISPR-dCas9-VPR technology reveals potent N-glycome alterations in colorectal cancer cells
    Article Snippet: .. The PCR product encoding the Puromycin marker was cloned into the pX330 vector after backbone linearization using two neighboring Sma I restriction sites (New England BioLabs) and InFusion cloning (Clontech, California, USA). .. Next, the Cas9 coding sequence was removed from the vector backbone by Nco I and Eco RI (New England BioLabs) digests.

    Article Title: Transcriptional activation of fucosyltransferase (FUT) genes using the CRISPR-dCas9-VPR technology reveals potent N-glycome alterations in colorectal cancer cells
    Article Snippet: .. The PCR product encoding the Puromycin marker was cloned into the pX330 vector after backbone linearization using two neighboring Sma I restriction sites (New England BioLabs) and InFusion cloning (Clontech, California, USA). .. Next, the Cas9 coding sequence was removed from the vector backbone by Nco I and Eco RI (New England BioLabs) digests.

    Article Title: Genome editing reveals a role for OCT4 in human embryogenesis
    Article Snippet: .. The sgRNA sequence from the correctly targeted px330 vector was amplified using the Q5 hot start high fidelity DNA polymerase (NEB; M0493) and the PCR product was in vitro transcribed using the MEGAshortscript T7 kit (ThermoFisher Scientific; AM1354) and purified using the Zymo RNA Clean & Concentrator columns (Zymo Research; R1017) The sgRNA and Cas9 mRNA (TriLink Biotechnologies; L61256) and recombinant Cas9 protein (Toolgen; TGEN CP1) were individually re-suspended in RNase-free water, aliquoted and stored at −80 °C until use. .. Prior to microinjection, the ribonucleoprotein complex was prepared by centrifuging the Cas9 protein for 1 min at 14,000 r.p.m. at 4 °C and transferring the supernatant to a fresh tube containing the sgRNA.

    Purification:

    Article Title: Genome editing reveals a role for OCT4 in human embryogenesis
    Article Snippet: .. The sgRNA sequence from the correctly targeted px330 vector was amplified using the Q5 hot start high fidelity DNA polymerase (NEB; M0493) and the PCR product was in vitro transcribed using the MEGAshortscript T7 kit (ThermoFisher Scientific; AM1354) and purified using the Zymo RNA Clean & Concentrator columns (Zymo Research; R1017) The sgRNA and Cas9 mRNA (TriLink Biotechnologies; L61256) and recombinant Cas9 protein (Toolgen; TGEN CP1) were individually re-suspended in RNase-free water, aliquoted and stored at −80 °C until use. .. Prior to microinjection, the ribonucleoprotein complex was prepared by centrifuging the Cas9 protein for 1 min at 14,000 r.p.m. at 4 °C and transferring the supernatant to a fresh tube containing the sgRNA.

    Marker:

    Article Title: Transcriptional activation of fucosyltransferase (FUT) genes using the CRISPR-dCas9-VPR technology reveals potent N-glycome alterations in colorectal cancer cells
    Article Snippet: .. The PCR product encoding the Puromycin marker was cloned into the pX330 vector after backbone linearization using two neighboring Sma I restriction sites (New England BioLabs) and InFusion cloning (Clontech, California, USA). .. Next, the Cas9 coding sequence was removed from the vector backbone by Nco I and Eco RI (New England BioLabs) digests.

    Article Title: Transcriptional activation of fucosyltransferase (FUT) genes using the CRISPR-dCas9-VPR technology reveals potent N-glycome alterations in colorectal cancer cells
    Article Snippet: .. The PCR product encoding the Puromycin marker was cloned into the pX330 vector after backbone linearization using two neighboring Sma I restriction sites (New England BioLabs) and InFusion cloning (Clontech, California, USA). .. Next, the Cas9 coding sequence was removed from the vector backbone by Nco I and Eco RI (New England BioLabs) digests.

    Sequencing:

    Article Title: Drug‐induced increase in lysobisphosphatidic acid reduces the cholesterol overload in Niemann–Pick type C cells and mice
    Article Snippet: .. The sequences were used to insert the target sequence into the pX330 vector using Golden Gate Assembly (New England Biolabs) and transfected into cells. .. Knockout clones were isolated by serial dilution and confirmed by RT–PCR, Western blotting and filipin staining.

    Article Title: Genome editing reveals a role for OCT4 in human embryogenesis
    Article Snippet: .. The sgRNA sequence from the correctly targeted px330 vector was amplified using the Q5 hot start high fidelity DNA polymerase (NEB; M0493) and the PCR product was in vitro transcribed using the MEGAshortscript T7 kit (ThermoFisher Scientific; AM1354) and purified using the Zymo RNA Clean & Concentrator columns (Zymo Research; R1017) The sgRNA and Cas9 mRNA (TriLink Biotechnologies; L61256) and recombinant Cas9 protein (Toolgen; TGEN CP1) were individually re-suspended in RNase-free water, aliquoted and stored at −80 °C until use. .. Prior to microinjection, the ribonucleoprotein complex was prepared by centrifuging the Cas9 protein for 1 min at 14,000 r.p.m. at 4 °C and transferring the supernatant to a fresh tube containing the sgRNA.

    Article Title: Cyclodextrin triggers MCOLN1-dependent endo-lysosome secretion in Niemann-Pick type C cells [S]
    Article Snippet: .. The sequences were used to insert the target sequence into the pX330 vector using Golden Gate Assembly (New England Biolabs) and transfected into cells. .. KO clones were isolated by serial dilution and confirmed by RT-PCR, Western blotting, and filipin staining.

    Recombinant:

    Article Title: Genome editing reveals a role for OCT4 in human embryogenesis
    Article Snippet: .. The sgRNA sequence from the correctly targeted px330 vector was amplified using the Q5 hot start high fidelity DNA polymerase (NEB; M0493) and the PCR product was in vitro transcribed using the MEGAshortscript T7 kit (ThermoFisher Scientific; AM1354) and purified using the Zymo RNA Clean & Concentrator columns (Zymo Research; R1017) The sgRNA and Cas9 mRNA (TriLink Biotechnologies; L61256) and recombinant Cas9 protein (Toolgen; TGEN CP1) were individually re-suspended in RNase-free water, aliquoted and stored at −80 °C until use. .. Prior to microinjection, the ribonucleoprotein complex was prepared by centrifuging the Cas9 protein for 1 min at 14,000 r.p.m. at 4 °C and transferring the supernatant to a fresh tube containing the sgRNA.

    Plasmid Preparation:

    Article Title: Transcriptional activation of fucosyltransferase (FUT) genes using the CRISPR-dCas9-VPR technology reveals potent N-glycome alterations in colorectal cancer cells
    Article Snippet: .. The PCR product encoding the Puromycin marker was cloned into the pX330 vector after backbone linearization using two neighboring Sma I restriction sites (New England BioLabs) and InFusion cloning (Clontech, California, USA). .. Next, the Cas9 coding sequence was removed from the vector backbone by Nco I and Eco RI (New England BioLabs) digests.

    Article Title: Rapid generation of gene-targeted EPS-derived mouse models through tetraploid complementation
    Article Snippet: .. The pX330 vector was digested by BbsI (NEB) for 16 h at 37 °C. .. The IL3 or IL6 gRNA-pX330 vector was constructed by ligating the gRNA annealed product and the pX330 digested product.

    Article Title: Drug‐induced increase in lysobisphosphatidic acid reduces the cholesterol overload in Niemann–Pick type C cells and mice
    Article Snippet: .. The sequences were used to insert the target sequence into the pX330 vector using Golden Gate Assembly (New England Biolabs) and transfected into cells. .. Knockout clones were isolated by serial dilution and confirmed by RT–PCR, Western blotting and filipin staining.

    Article Title: Transcriptional activation of fucosyltransferase (FUT) genes using the CRISPR-dCas9-VPR technology reveals potent N-glycome alterations in colorectal cancer cells
    Article Snippet: .. The PCR product encoding the Puromycin marker was cloned into the pX330 vector after backbone linearization using two neighboring Sma I restriction sites (New England BioLabs) and InFusion cloning (Clontech, California, USA). .. Next, the Cas9 coding sequence was removed from the vector backbone by Nco I and Eco RI (New England BioLabs) digests.

    Article Title: Generation of WNK1 knockout cell lines by CRISPR/Cas-mediated genome editing
    Article Snippet: .. The annealed oligo was ligated into the BbsI -digested pX330 vector using 5 μl of 2× QuickLigation Buffer and 1 μl of QuickLigase (New England Biolabs). .. The ligation mixture was treated with PlasmidSafe exonuclease (Epicentre) and transformed in OneShot chemically competent Stbl3 cells (Life Technologies).

    Article Title: Genome editing reveals a role for OCT4 in human embryogenesis
    Article Snippet: .. The sgRNA sequence from the correctly targeted px330 vector was amplified using the Q5 hot start high fidelity DNA polymerase (NEB; M0493) and the PCR product was in vitro transcribed using the MEGAshortscript T7 kit (ThermoFisher Scientific; AM1354) and purified using the Zymo RNA Clean & Concentrator columns (Zymo Research; R1017) The sgRNA and Cas9 mRNA (TriLink Biotechnologies; L61256) and recombinant Cas9 protein (Toolgen; TGEN CP1) were individually re-suspended in RNase-free water, aliquoted and stored at −80 °C until use. .. Prior to microinjection, the ribonucleoprotein complex was prepared by centrifuging the Cas9 protein for 1 min at 14,000 r.p.m. at 4 °C and transferring the supernatant to a fresh tube containing the sgRNA.

    Article Title: Cyclodextrin triggers MCOLN1-dependent endo-lysosome secretion in Niemann-Pick type C cells [S]
    Article Snippet: .. The sequences were used to insert the target sequence into the pX330 vector using Golden Gate Assembly (New England Biolabs) and transfected into cells. .. KO clones were isolated by serial dilution and confirmed by RT-PCR, Western blotting, and filipin staining.

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  • Team
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  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 91
    New England Biolabs px330 vector
    Generation and validation of WNK1 knockout (KO) cell lines. A : mismatch-specific endonuclease assay. Genomic PCR (gPCR) products spanning exon 1 of WNK1 were amplified from the template of a heterogeneous population of HEK293T cells transfected with <t>px330-WNK1</t>
    Px330 Vector, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/px330 vector/product/New England Biolabs
    Average 91 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    px330 vector - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

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    Generation and validation of WNK1 knockout (KO) cell lines. A : mismatch-specific endonuclease assay. Genomic PCR (gPCR) products spanning exon 1 of WNK1 were amplified from the template of a heterogeneous population of HEK293T cells transfected with px330-WNK1

    Journal: American Journal of Physiology - Renal Physiology

    Article Title: Generation of WNK1 knockout cell lines by CRISPR/Cas-mediated genome editing

    doi: 10.1152/ajprenal.00612.2014

    Figure Lengend Snippet: Generation and validation of WNK1 knockout (KO) cell lines. A : mismatch-specific endonuclease assay. Genomic PCR (gPCR) products spanning exon 1 of WNK1 were amplified from the template of a heterogeneous population of HEK293T cells transfected with px330-WNK1

    Article Snippet: The annealed oligo was ligated into the BbsI -digested pX330 vector using 5 μl of 2× QuickLigation Buffer and 1 μl of QuickLigase (New England Biolabs).

    Techniques: Knock-Out, Polymerase Chain Reaction, Amplification, Transfection