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fncas12a and ascas12a expression constructs  (GenScript corporation)

 
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    GenScript corporation fncas12a and ascas12a expression constructs
    Transformation of <t>FnCas12a-mediated</t> targeting plasmids in Bacillus smithii ET 138. (A) Schematic representation of the basic pFnCas12a plasmid. The fncas12a gene was introduced in the pNW33n vector backbone. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Results of the OD600 measurements from cultures of B. smithii transformed with pFnCas12a_NT compared to an empty pNW33n plasmid. (C) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a expression and targeting efficiency at different temperatures; detailed description of the protocol can be found in the Methods section. (D) Results of the OD600 measurements from cultures of B. smithii transformed with targeting plasmids (pFnCas12a_Sp1/2/3) normalized to that of cultures harboring a non-targeting (NT) plasmid (pFnCas12a_NT). Average values of three replicates are shown, with error bars representing SD.
    Fncas12a And Ascas12a Expression Constructs, supplied by GenScript corporation, 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/result/fncas12a and ascas12a expression constructs/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    fncas12a and ascas12a expression constructs - by Bioz Stars, 2026-03
    90/100 stars

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    1) Product Images from "Development of a Cas12a-Based Genome Editing Tool for Moderate Thermophiles"

    Article Title: Development of a Cas12a-Based Genome Editing Tool for Moderate Thermophiles

    Journal: The CRISPR Journal

    doi: 10.1089/crispr.2020.0086

    Transformation of FnCas12a-mediated targeting plasmids in Bacillus smithii ET 138. (A) Schematic representation of the basic pFnCas12a plasmid. The fncas12a gene was introduced in the pNW33n vector backbone. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Results of the OD600 measurements from cultures of B. smithii transformed with pFnCas12a_NT compared to an empty pNW33n plasmid. (C) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a expression and targeting efficiency at different temperatures; detailed description of the protocol can be found in the Methods section. (D) Results of the OD600 measurements from cultures of B. smithii transformed with targeting plasmids (pFnCas12a_Sp1/2/3) normalized to that of cultures harboring a non-targeting (NT) plasmid (pFnCas12a_NT). Average values of three replicates are shown, with error bars representing SD.
    Figure Legend Snippet: Transformation of FnCas12a-mediated targeting plasmids in Bacillus smithii ET 138. (A) Schematic representation of the basic pFnCas12a plasmid. The fncas12a gene was introduced in the pNW33n vector backbone. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Results of the OD600 measurements from cultures of B. smithii transformed with pFnCas12a_NT compared to an empty pNW33n plasmid. (C) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a expression and targeting efficiency at different temperatures; detailed description of the protocol can be found in the Methods section. (D) Results of the OD600 measurements from cultures of B. smithii transformed with targeting plasmids (pFnCas12a_Sp1/2/3) normalized to that of cultures harboring a non-targeting (NT) plasmid (pFnCas12a_NT). Average values of three replicates are shown, with error bars representing SD.

    Techniques Used: Transformation Assay, Plasmid Preparation, Expressing, Marker

    Transformation of HR-FnCas12a mediated editing plasmids in B. smithii ET 138. (A) Schematic representation of the pFnCas12a_Δgene-of-interest (goi)-HR construct. The fncas12a gene was introduced to the pNW33n vector backbone. Homologous recombination (HR) flanks were introduced upstream fncas12a gene and encompassed the 1 kb upstream and 1 kb downstream region of the goi in the B. smithii genome. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a editing efficiency; detailed description of the protocol can be found in the Methods section. (C) Agarose gel electrophoresis showing the results from polymerase chain reaction on the genomic DNA of B. smithii cultures transformed with pFnCas12a_ΔpyrF-HR_Sp1 (1), pFnCas12a_ΔpyrF-HR_Sp2 (2), and pFnCas12a_ΔpyrF-HR_NT (NT) in two different selection media (TVMYxgu and LB2xgu). The last two lanes are the negative (wild type) and positive (ΔpyrF) controls that correspond to DNA fragments 2.9 and 2.2 kb long, respectively. (D) Representative image of the sequence verification of the desired pyrF gene deletion by Sanger sequencing.
    Figure Legend Snippet: Transformation of HR-FnCas12a mediated editing plasmids in B. smithii ET 138. (A) Schematic representation of the pFnCas12a_Δgene-of-interest (goi)-HR construct. The fncas12a gene was introduced to the pNW33n vector backbone. Homologous recombination (HR) flanks were introduced upstream fncas12a gene and encompassed the 1 kb upstream and 1 kb downstream region of the goi in the B. smithii genome. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a editing efficiency; detailed description of the protocol can be found in the Methods section. (C) Agarose gel electrophoresis showing the results from polymerase chain reaction on the genomic DNA of B. smithii cultures transformed with pFnCas12a_ΔpyrF-HR_Sp1 (1), pFnCas12a_ΔpyrF-HR_Sp2 (2), and pFnCas12a_ΔpyrF-HR_NT (NT) in two different selection media (TVMYxgu and LB2xgu). The last two lanes are the negative (wild type) and positive (ΔpyrF) controls that correspond to DNA fragments 2.9 and 2.2 kb long, respectively. (D) Representative image of the sequence verification of the desired pyrF gene deletion by Sanger sequencing.

    Techniques Used: Transformation Assay, Construct, Plasmid Preparation, Homologous Recombination, Expressing, Marker, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Selection, Sequencing



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    fncas12a and ascas12a expression constructs  (GenScript corporation)
    90
    GenScript corporation fncas12a and ascas12a expression constructs
    Transformation of <t>FnCas12a-mediated</t> targeting plasmids in Bacillus smithii ET 138. (A) Schematic representation of the basic pFnCas12a plasmid. The fncas12a gene was introduced in the pNW33n vector backbone. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Results of the OD600 measurements from cultures of B. smithii transformed with pFnCas12a_NT compared to an empty pNW33n plasmid. (C) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a expression and targeting efficiency at different temperatures; detailed description of the protocol can be found in the Methods section. (D) Results of the OD600 measurements from cultures of B. smithii transformed with targeting plasmids (pFnCas12a_Sp1/2/3) normalized to that of cultures harboring a non-targeting (NT) plasmid (pFnCas12a_NT). Average values of three replicates are shown, with error bars representing SD.
    Fncas12a And Ascas12a Expression Constructs, supplied by GenScript corporation, 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/result/fncas12a and ascas12a expression constructs/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    fncas12a and ascas12a expression constructs - by Bioz Stars, 2026-03
    90/100 stars
    		  Buy from Supplier

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    Transformation of FnCas12a-mediated targeting plasmids in Bacillus smithii ET 138. (A) Schematic representation of the basic pFnCas12a plasmid. The fncas12a gene was introduced in the pNW33n vector backbone. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Results of the OD600 measurements from cultures of B. smithii transformed with pFnCas12a_NT compared to an empty pNW33n plasmid. (C) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a expression and targeting efficiency at different temperatures; detailed description of the protocol can be found in the Methods section. (D) Results of the OD600 measurements from cultures of B. smithii transformed with targeting plasmids (pFnCas12a_Sp1/2/3) normalized to that of cultures harboring a non-targeting (NT) plasmid (pFnCas12a_NT). Average values of three replicates are shown, with error bars representing SD.

    Journal: The CRISPR Journal

    Article Title: Development of a Cas12a-Based Genome Editing Tool for Moderate Thermophiles

    doi: 10.1089/crispr.2020.0086

    Figure Lengend Snippet: Transformation of FnCas12a-mediated targeting plasmids in Bacillus smithii ET 138. (A) Schematic representation of the basic pFnCas12a plasmid. The fncas12a gene was introduced in the pNW33n vector backbone. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Results of the OD600 measurements from cultures of B. smithii transformed with pFnCas12a_NT compared to an empty pNW33n plasmid. (C) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a expression and targeting efficiency at different temperatures; detailed description of the protocol can be found in the Methods section. (D) Results of the OD600 measurements from cultures of B. smithii transformed with targeting plasmids (pFnCas12a_Sp1/2/3) normalized to that of cultures harboring a non-targeting (NT) plasmid (pFnCas12a_NT). Average values of three replicates are shown, with error bars representing SD.

    Article Snippet: 21 The FnCas12a and AsCas12a expression constructs were synthesized by GenScript.

    Techniques: Transformation Assay, Plasmid Preparation, Expressing, Marker

    Transformation of HR-FnCas12a mediated editing plasmids in B. smithii ET 138. (A) Schematic representation of the pFnCas12a_Δgene-of-interest (goi)-HR construct. The fncas12a gene was introduced to the pNW33n vector backbone. Homologous recombination (HR) flanks were introduced upstream fncas12a gene and encompassed the 1 kb upstream and 1 kb downstream region of the goi in the B. smithii genome. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a editing efficiency; detailed description of the protocol can be found in the Methods section. (C) Agarose gel electrophoresis showing the results from polymerase chain reaction on the genomic DNA of B. smithii cultures transformed with pFnCas12a_ΔpyrF-HR_Sp1 (1), pFnCas12a_ΔpyrF-HR_Sp2 (2), and pFnCas12a_ΔpyrF-HR_NT (NT) in two different selection media (TVMYxgu and LB2xgu). The last two lanes are the negative (wild type) and positive (ΔpyrF) controls that correspond to DNA fragments 2.9 and 2.2 kb long, respectively. (D) Representative image of the sequence verification of the desired pyrF gene deletion by Sanger sequencing.

    Journal: The CRISPR Journal

    Article Title: Development of a Cas12a-Based Genome Editing Tool for Moderate Thermophiles

    doi: 10.1089/crispr.2020.0086

    Figure Lengend Snippet: Transformation of HR-FnCas12a mediated editing plasmids in B. smithii ET 138. (A) Schematic representation of the pFnCas12a_Δgene-of-interest (goi)-HR construct. The fncas12a gene was introduced to the pNW33n vector backbone. Homologous recombination (HR) flanks were introduced upstream fncas12a gene and encompassed the 1 kb upstream and 1 kb downstream region of the goi in the B. smithii genome. A crRNA-expressing module was introduced downstream the fncas12a gene. Other elements on the plasmid are origin of replication (ori), replication protein (repB), and chloramphenicol-resistance marker (CmR). (B) Sequential transfer scheme of B. smithii cultures to evaluate FnCas12a editing efficiency; detailed description of the protocol can be found in the Methods section. (C) Agarose gel electrophoresis showing the results from polymerase chain reaction on the genomic DNA of B. smithii cultures transformed with pFnCas12a_ΔpyrF-HR_Sp1 (1), pFnCas12a_ΔpyrF-HR_Sp2 (2), and pFnCas12a_ΔpyrF-HR_NT (NT) in two different selection media (TVMYxgu and LB2xgu). The last two lanes are the negative (wild type) and positive (ΔpyrF) controls that correspond to DNA fragments 2.9 and 2.2 kb long, respectively. (D) Representative image of the sequence verification of the desired pyrF gene deletion by Sanger sequencing.

    Article Snippet: 21 The FnCas12a and AsCas12a expression constructs were synthesized by GenScript.

    Techniques: Transformation Assay, Construct, Plasmid Preparation, Homologous Recombination, Expressing, Marker, Agarose Gel Electrophoresis, Polymerase Chain Reaction, Selection, Sequencing