px330 vector  (New England Biolabs)


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    Structured Review

    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: 90/100, based on 2169 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/px330 vector/product/New England Biolabs
    Average 90 stars, based on 2169 article reviews
    Price from $9.99 to $1999.99
    px330 vector - by Bioz Stars, 2021-01
    90/100 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

    In Vitro:

    Article Title: A Simple and Cost-Effective Approach for In Vitro Production of Sliced siRNAs as Potent Triggers for RNAi
    Article Snippet: .. The protocol was modified as follows when CIP or T4 PNK treatment is necessary: (1) for CIP treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of CIP, 4 μL of 10× CutSmart buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min; and (2) for T4 PNK treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of T4 PNK, 4 μL of 10× T4 PNK buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min. All T7 in vitro transcription products were purified by Micro Bio-Spin P-30 Gel Columns, Tris Buffer, from Bio-Rad. ..

    Modification:

    Article Title: A Simple and Cost-Effective Approach for In Vitro Production of Sliced siRNAs as Potent Triggers for RNAi
    Article Snippet: .. The protocol was modified as follows when CIP or T4 PNK treatment is necessary: (1) for CIP treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of CIP, 4 μL of 10× CutSmart buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min; and (2) for T4 PNK treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of T4 PNK, 4 μL of 10× T4 PNK buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min. All T7 in vitro transcription products were purified by Micro Bio-Spin P-30 Gel Columns, Tris Buffer, from Bio-Rad. ..

    Ethanol Precipitation:

    Article Title: tRNA-derived fragments and microRNAs in the maternal-fetal interface of a mouse maternal-immune-activation autism model
    Article Snippet: .. T4 PNK treatment and RT-PCR1 μg total RNA was treated with T4 PNK (NEB) in the presence of 10 mM ATP at 37 °C for 40 minutes and then cleaned up by ethanol precipitation. ..

    Labeling:

    Article Title: Deep small RNA sequencing from the nematode Ascaris reveals conservation, functional diversification, and novel developmental profiles
    Article Snippet: .. Total RNA was isolated using TRIzol (Invitrogen), and small RNA samples were 5′ labeled by first treating with calf alkaline phosphatase (Roche) followed by phosphorylation with T4 polynucleotide kinase (NEB) and 32 P-γ-ATP. .. Small RNA samples were 3′ labeled using T4 RNA ligase (NEB) and 32 P-pCp as described by the manufacturer.

    Article Title: Kaposi's Sarcoma-Associated Herpesvirus Rta Tetramers Make High-Affinity Interactions with Repetitive DNA Elements in the Mta Promoter To Stimulate DNA Binding of RBP-Jk/CSL ▿Kaposi's Sarcoma-Associated Herpesvirus Rta Tetramers Make High-Affinity Interactions with Repetitive DNA Elements in the Mta Promoter To Stimulate DNA Binding of RBP-Jk/CSL ▿ †
    Article Snippet: .. Five hundred nanograms of probe was end labeled using 10 units T4 polynucleotide kinase (NEB) and 20 μCi of [γ32 -P]dATP (PerkinElmer) in a total volume of 50 μl for 1 h at 37°C. .. Free nucleotide was removed by Sephadex purification (USA Scientific) per the manufacturer's suggestions.

    Purification:

    Article Title: A Simple and Cost-Effective Approach for In Vitro Production of Sliced siRNAs as Potent Triggers for RNAi
    Article Snippet: .. The protocol was modified as follows when CIP or T4 PNK treatment is necessary: (1) for CIP treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of CIP, 4 μL of 10× CutSmart buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min; and (2) for T4 PNK treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of T4 PNK, 4 μL of 10× T4 PNK buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min. All T7 in vitro transcription products were purified by Micro Bio-Spin P-30 Gel Columns, Tris Buffer, from Bio-Rad. ..

    Article Title: CspA regulation of Staphylococcus aureus carotenoid levels and σB activity is controlled by YjbH and Spx.
    Article Snippet: .. To decipher interactions between proteins and the cspA promoter, a PCR product consisting of −300 to +1 of the cspA promoter was first end-labeled with γ−32P-ATP (Perkin Elmer) using T4 polynucleotide kinase (NEB) and purified by MicroSpin G-50 columns (GE). .. His6 -Spx and/or αCTD were pre-incubated for 10 min at 25oC in 18 μL of binding buffer (20 mM Tris-HCl pH 7.8, 50 mM NaCl, 5 mM MgCl2, 10% glycerol) containing 0.5 μg of calf thymus DNA.

    Produced:

    Article Title: PARP3 is a sensor of nicked nucleosomes and monoribosylates histone H2BGlu2
    Article Snippet: .. DNA from the MNase concentration that produced the greatest SSB/DSB ratio (0.015 U) was mock-treated or treated with T4 PNK in the presence of 2 mM ATP and 10U T4 PNK enzyme (wild-type or 3′-phosphatase dead; New England Biolabs). .. The synthetic oligonucleotide sequences (MWG or Eurogentec) employed to generate duplex substrates are listed in .

    Concentration Assay:

    Article Title: PARP3 is a sensor of nicked nucleosomes and monoribosylates histone H2BGlu2
    Article Snippet: .. DNA from the MNase concentration that produced the greatest SSB/DSB ratio (0.015 U) was mock-treated or treated with T4 PNK in the presence of 2 mM ATP and 10U T4 PNK enzyme (wild-type or 3′-phosphatase dead; New England Biolabs). .. The synthetic oligonucleotide sequences (MWG or Eurogentec) employed to generate duplex substrates are listed in .

    Incubation:

    Article Title: A Simple and Cost-Effective Approach for In Vitro Production of Sliced siRNAs as Potent Triggers for RNAi
    Article Snippet: .. The protocol was modified as follows when CIP or T4 PNK treatment is necessary: (1) for CIP treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of CIP, 4 μL of 10× CutSmart buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min; and (2) for T4 PNK treatment, in 20 μL of products from one in vitro transcription reaction before DNase treatment, we added 1 μL of DNase (supplied with T7 Transcription Kit), 1 μL of T4 PNK, 4 μL of 10× T4 PNK buffer (NEB), and water to total volume of 40 μL, and incubated at 37°C for 15 min. All T7 in vitro transcription products were purified by Micro Bio-Spin P-30 Gel Columns, Tris Buffer, from Bio-Rad. ..

    other:

    Article Title: Distinct and overlapping roles for two Dicer-like proteins in the RNA interference pathways of the ancient eukaryote Trypanosoma brucei
    Article Snippet: Small RNAs were size-selected ( ) and treated with a variety of enzymes under manufacturer-recommended conditions: Calf intestinal alkaline phosphatase (CIP; Amersham) and T4 polynucleotide kinase (T4 PNK; New England Biolabs) assays were carried out for 1 h at 37 °C, and Terminator exonuclease (Epicentre) was added for 1 h at 30 °C.

    Isolation:

    Article Title: Deep small RNA sequencing from the nematode Ascaris reveals conservation, functional diversification, and novel developmental profiles
    Article Snippet: .. Total RNA was isolated using TRIzol (Invitrogen), and small RNA samples were 5′ labeled by first treating with calf alkaline phosphatase (Roche) followed by phosphorylation with T4 polynucleotide kinase (NEB) and 32 P-γ-ATP. .. Small RNA samples were 3′ labeled using T4 RNA ligase (NEB) and 32 P-pCp as described by the manufacturer.

    Polymerase Chain Reaction:

    Article Title: CspA regulation of Staphylococcus aureus carotenoid levels and σB activity is controlled by YjbH and Spx.
    Article Snippet: .. To decipher interactions between proteins and the cspA promoter, a PCR product consisting of −300 to +1 of the cspA promoter was first end-labeled with γ−32P-ATP (Perkin Elmer) using T4 polynucleotide kinase (NEB) and purified by MicroSpin G-50 columns (GE). .. His6 -Spx and/or αCTD were pre-incubated for 10 min at 25oC in 18 μL of binding buffer (20 mM Tris-HCl pH 7.8, 50 mM NaCl, 5 mM MgCl2, 10% glycerol) containing 0.5 μg of calf thymus DNA.

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    New England Biolabs quickligase kit
    Quickligase Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quickligase kit/product/New England Biolabs
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    Price from $9.99 to $1999.99
    quickligase kit - by Bioz Stars, 2021-01
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