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plasmid pgem1  (Promega)

 
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    Promega plasmid pgem1
    Plasmid Pgem1, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/plasmid+pgem1/pgem+t+easy/pm39970206-235-5-15
    Average 90 stars, based on 1 article reviews
    plasmid pgem1 - by Bioz Stars, 2026-10
    90/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: Sequence-dependent scale for translocon-mediated insertion of interfacial helices in membranes.
    Article Snippet: .. All enzymes as well as plasmid pGEM1 and the TnT coupled transcription/translation system were from Promega (Madison, WI). ..

    Article Title: Membrane insertion and topology of the translocon-associated protein (TRAP) gamma subunit.
    Article Snippet: .. All enzymes, as well as plasmid pGEM1, RiboMAX SP6 RNA polymerase system and rabbit reticulocyte lysate were from Promega (Madison, WI). ..

    Article Title: The C-terminal Domains of Apoptotic BH3-only Proteins Mediate Their Insertion into Distinct Biological Membranes
    Article Snippet: .. All enzymes, as well as plasmid pGEM1, and the T n T SP6 Quick Coupled System were from Promega (Madison, WI). ..

    Article Title: Murine astrotactins 1 and 2 have a similar membrane topology and mature via endoproteolytic cleavage catalyzed by a signal peptidase
    Article Snippet: Enzymes and chemicals Unless otherwise stated, all chemicals were from Sigma-Aldrich (St. Louis, MO). .. Plasmid pGEM1, the T n T® Quick Coupled Transcription/Translation System, the rabbit reticulocyte lysate system, and deoxynucleotides were from Promega (Madison, WI). .. [ 35 S]Met was from PerkinElmer Life Sciences.

    Article Title: Membrane integration and topology of RIFIN and STEVOR proteins of the Plasmodium falciparum parasite.
    Article Snippet: The malarial parasite Plasmodium exports its own proteins to the cell surfaces of red blood cells during infection.. Examples of exported proteins include members of the repetitive interspersed family (RIFIN) and sub-telomeric variable open reading frame (STEVOR) family of proteins from Plasmodium falciparum.. The presence of these parasite-derived proteins on surfaces of infected red blood cells triggers the adhesion of infected cells to uninfected cells (rosetting), and to the vascular endothelium potentially obstructing blood flow.

    Article Title: Murine astrotactins 1 and 2 have a similar membrane topology and mature via endoproteolytic cleavage catalyzed by a signal peptidase
    Article Snippet: Experimental procedures Enzymes and chemicals – Unless otherwise stated, all chemicals were from Sigma-Aldrich (St. Louis, MO, US). .. Plasmid pGEM1, TNT® Quick Coupled transcription/translation system, Rabbit Reticulocyte lysate system and deoxynucleotides were from Promega (Madison, WI, US). .. [35S]-Met was from PerkinElmer (Boston MA, US).

    other:

    Article Title: Anti-IL13 antibodies and uses thereof
    Article Snippet: Examples of useful expression vectors for prokaryotic host cells include those derived from commercially available plasmids such as the pKK223-3 (Pharmacia Fine Chemicals, Uppsala, Sweden), pGEM1 (Promega Biotec, Madison, Wis., USA), and the pET (Novagen, Madison, Wis., USA) and pRSET (Invitrogen Corporation, Carlsbad, Calif., USA) series of vectors (Studier, F. W., J. Mol.



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    pgem1 plasmid  (New England Biolabs)
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    New England Biolabs pgem1 plasmid
    General workflow for CRISPR‐based site‐specific mutagenesis of multi‐allelic genes in cancer cell lines. (A) Three plasmids used for mutagenesis. The plasmid expressing spCas9 and the sgRNA (pX459) and the repair template plasmid containing the homology arms to the intended target site as well as the desired nucleotide substitutions for mutagenesis <t>(pGEM1)</t> are transfected into the cells. A third plasmid, containing a larger fragment of the endogenous locus sequence plus the desired nucleotide substitutions for mutagenesis, is used as a positive control for the ASP genotyping strategy. (B) Transfections, antibiotic selection, colony picking, and genomic DNA extraction for genotyping of potential KI clones are discussed in the text. (C) ASP is used as a screening strategy to identify clones that have successfully integrated the repair template sequence with desired nucleotide substitutions (KI). (D) Western blotting and mass spectrometry (LC‐MS/MS) is used to confirm that KI clones successfully produce full‐length KI proteins. (E, F) Allele frequency estimation is performed using TOPO cloning‐based screening of genomic DNA extracted from individual KI clones and/or the use of next‐generation sequencing approaches such as RNA‐sequencing to measure the abundance of RNA transcripts that contain the desired KI mutations. The details of each step are discussed throughout the text.
    Pgem1 Plasmid, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/plasmid+pgem1/Q5+Site-Directed+Mutagenesis+Kit/pmc08167868-88-4-13
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    plasmid pgem1  (Promega)
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    Promega plasmid pgem1
    General workflow for CRISPR‐based site‐specific mutagenesis of multi‐allelic genes in cancer cell lines. (A) Three plasmids used for mutagenesis. The plasmid expressing spCas9 and the sgRNA (pX459) and the repair template plasmid containing the homology arms to the intended target site as well as the desired nucleotide substitutions for mutagenesis <t>(pGEM1)</t> are transfected into the cells. A third plasmid, containing a larger fragment of the endogenous locus sequence plus the desired nucleotide substitutions for mutagenesis, is used as a positive control for the ASP genotyping strategy. (B) Transfections, antibiotic selection, colony picking, and genomic DNA extraction for genotyping of potential KI clones are discussed in the text. (C) ASP is used as a screening strategy to identify clones that have successfully integrated the repair template sequence with desired nucleotide substitutions (KI). (D) Western blotting and mass spectrometry (LC‐MS/MS) is used to confirm that KI clones successfully produce full‐length KI proteins. (E, F) Allele frequency estimation is performed using TOPO cloning‐based screening of genomic DNA extracted from individual KI clones and/or the use of next‐generation sequencing approaches such as RNA‐sequencing to measure the abundance of RNA transcripts that contain the desired KI mutations. The details of each step are discussed throughout the text.
    Plasmid Pgem1, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/plasmid+pgem1/pgem+t+easy/pm39970206-235-5-15
    Average 90 stars, based on 1 article reviews
    plasmid pgem1 - by Bioz Stars, 2026-10
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    		  Buy from Supplier
    pgem1-derived plasmid  (Promega)
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    Promega pgem1-derived plasmid
    General workflow for CRISPR‐based site‐specific mutagenesis of multi‐allelic genes in cancer cell lines. (A) Three plasmids used for mutagenesis. The plasmid expressing spCas9 and the sgRNA (pX459) and the repair template plasmid containing the homology arms to the intended target site as well as the desired nucleotide substitutions for mutagenesis <t>(pGEM1)</t> are transfected into the cells. A third plasmid, containing a larger fragment of the endogenous locus sequence plus the desired nucleotide substitutions for mutagenesis, is used as a positive control for the ASP genotyping strategy. (B) Transfections, antibiotic selection, colony picking, and genomic DNA extraction for genotyping of potential KI clones are discussed in the text. (C) ASP is used as a screening strategy to identify clones that have successfully integrated the repair template sequence with desired nucleotide substitutions (KI). (D) Western blotting and mass spectrometry (LC‐MS/MS) is used to confirm that KI clones successfully produce full‐length KI proteins. (E, F) Allele frequency estimation is performed using TOPO cloning‐based screening of genomic DNA extracted from individual KI clones and/or the use of next‐generation sequencing approaches such as RNA‐sequencing to measure the abundance of RNA transcripts that contain the desired KI mutations. The details of each step are discussed throughout the text.
    Pgem1 Derived Plasmid, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/plasmid+pgem1/pgem+t+easy/pmc06964287-51-10-24
    Average 90 stars, based on 1 article reviews
    pgem1-derived plasmid - by Bioz Stars, 2026-10
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    pgem1 plasmid  (Promega)
    90
    Promega pgem1 plasmid
    General workflow for CRISPR‐based site‐specific mutagenesis of multi‐allelic genes in cancer cell lines. (A) Three plasmids used for mutagenesis. The plasmid expressing spCas9 and the sgRNA (pX459) and the repair template plasmid containing the homology arms to the intended target site as well as the desired nucleotide substitutions for mutagenesis <t>(pGEM1)</t> are transfected into the cells. A third plasmid, containing a larger fragment of the endogenous locus sequence plus the desired nucleotide substitutions for mutagenesis, is used as a positive control for the ASP genotyping strategy. (B) Transfections, antibiotic selection, colony picking, and genomic DNA extraction for genotyping of potential KI clones are discussed in the text. (C) ASP is used as a screening strategy to identify clones that have successfully integrated the repair template sequence with desired nucleotide substitutions (KI). (D) Western blotting and mass spectrometry (LC‐MS/MS) is used to confirm that KI clones successfully produce full‐length KI proteins. (E, F) Allele frequency estimation is performed using TOPO cloning‐based screening of genomic DNA extracted from individual KI clones and/or the use of next‐generation sequencing approaches such as RNA‐sequencing to measure the abundance of RNA transcripts that contain the desired KI mutations. The details of each step are discussed throughout the text.
    Pgem1 Plasmid, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/plasmid+pgem1/pgem+t+easy/10__1074_slash_jbc__m116__740985-339-1-15
    Average 90 stars, based on 1 article reviews
    pgem1 plasmid - by Bioz Stars, 2026-10
    90/100 stars
    		  Buy from Supplier

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    General workflow for CRISPR‐based site‐specific mutagenesis of multi‐allelic genes in cancer cell lines. (A) Three plasmids used for mutagenesis. The plasmid expressing spCas9 and the sgRNA (pX459) and the repair template plasmid containing the homology arms to the intended target site as well as the desired nucleotide substitutions for mutagenesis (pGEM1) are transfected into the cells. A third plasmid, containing a larger fragment of the endogenous locus sequence plus the desired nucleotide substitutions for mutagenesis, is used as a positive control for the ASP genotyping strategy. (B) Transfections, antibiotic selection, colony picking, and genomic DNA extraction for genotyping of potential KI clones are discussed in the text. (C) ASP is used as a screening strategy to identify clones that have successfully integrated the repair template sequence with desired nucleotide substitutions (KI). (D) Western blotting and mass spectrometry (LC‐MS/MS) is used to confirm that KI clones successfully produce full‐length KI proteins. (E, F) Allele frequency estimation is performed using TOPO cloning‐based screening of genomic DNA extracted from individual KI clones and/or the use of next‐generation sequencing approaches such as RNA‐sequencing to measure the abundance of RNA transcripts that contain the desired KI mutations. The details of each step are discussed throughout the text.

    Journal: FEBS Open Bio

    Article Title: CRISPR‐based knock‐in mutagenesis of the pioneer transcription factor FOXA1: optimization of strategies for multi‐allelic proteins in cancer cells

    doi: 10.1002/2211-5463.13139

    Figure Lengend Snippet: General workflow for CRISPR‐based site‐specific mutagenesis of multi‐allelic genes in cancer cell lines. (A) Three plasmids used for mutagenesis. The plasmid expressing spCas9 and the sgRNA (pX459) and the repair template plasmid containing the homology arms to the intended target site as well as the desired nucleotide substitutions for mutagenesis (pGEM1) are transfected into the cells. A third plasmid, containing a larger fragment of the endogenous locus sequence plus the desired nucleotide substitutions for mutagenesis, is used as a positive control for the ASP genotyping strategy. (B) Transfections, antibiotic selection, colony picking, and genomic DNA extraction for genotyping of potential KI clones are discussed in the text. (C) ASP is used as a screening strategy to identify clones that have successfully integrated the repair template sequence with desired nucleotide substitutions (KI). (D) Western blotting and mass spectrometry (LC‐MS/MS) is used to confirm that KI clones successfully produce full‐length KI proteins. (E, F) Allele frequency estimation is performed using TOPO cloning‐based screening of genomic DNA extracted from individual KI clones and/or the use of next‐generation sequencing approaches such as RNA‐sequencing to measure the abundance of RNA transcripts that contain the desired KI mutations. The details of each step are discussed throughout the text.

    Article Snippet: Once cloned into the pGEM1 plasmid, we used the Q5 site‐directed mutagenesis Kit (NEB Cat# E0554S) as per the manufacturer's instructions to introduce the specific nucleic acid substitutions needed to create the repair template sequence shown in Fig. .

    Techniques: CRISPR, Mutagenesis, Plasmid Preparation, Expressing, Transfection, Sequencing, Positive Control, Selection, DNA Extraction, Clone Assay, Western Blot, Mass Spectrometry, Liquid Chromatography with Mass Spectroscopy, Cloning, Next-Generation Sequencing, RNA Sequencing

    Repair template plasmid and PCR screening positive control plasmid preparation. The human FOXA1 endogenous locus is shown at the top. Two different plasmids were generated, the repair template plasmid and an ASP positive control plasmid to be used during the screening of potential KI clones. The repair template plasmid was built on the pGEM1 backbone and contains the FOXA1 repair template sequence (shown in Fig. ) along with FOXA1 homology arms of 600 bp upstream and 1200 bp downstream of K295. The ASP ‐positive control plasmid was intentionally designed to contain more of the endogenous sequence of FOXA1 as well as the repair template sequence to be able to test our ASP primers. The forward primer only hybridizes to this plasmid and not the repair template plasmid thus allowing us to detect successful KI editing within the endogenous locus of FOXA1 and not random integration of the repair plasmid.

    Journal: FEBS Open Bio

    Article Title: CRISPR‐based knock‐in mutagenesis of the pioneer transcription factor FOXA1: optimization of strategies for multi‐allelic proteins in cancer cells

    doi: 10.1002/2211-5463.13139

    Figure Lengend Snippet: Repair template plasmid and PCR screening positive control plasmid preparation. The human FOXA1 endogenous locus is shown at the top. Two different plasmids were generated, the repair template plasmid and an ASP positive control plasmid to be used during the screening of potential KI clones. The repair template plasmid was built on the pGEM1 backbone and contains the FOXA1 repair template sequence (shown in Fig. ) along with FOXA1 homology arms of 600 bp upstream and 1200 bp downstream of K295. The ASP ‐positive control plasmid was intentionally designed to contain more of the endogenous sequence of FOXA1 as well as the repair template sequence to be able to test our ASP primers. The forward primer only hybridizes to this plasmid and not the repair template plasmid thus allowing us to detect successful KI editing within the endogenous locus of FOXA1 and not random integration of the repair plasmid.

    Article Snippet: Once cloned into the pGEM1 plasmid, we used the Q5 site‐directed mutagenesis Kit (NEB Cat# E0554S) as per the manufacturer's instructions to introduce the specific nucleic acid substitutions needed to create the repair template sequence shown in Fig. .

    Techniques: Plasmid Preparation, Positive Control, Generated, Clone Assay, Sequencing