restriction enzyme bpuei  (New England Biolabs)


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    Name:
    BpuEI
    Description:
    BpuEI 500 units
    Catalog Number:
    r0633s
    Price:
    69
    Size:
    500 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs restriction enzyme bpuei
    BpuEI
    BpuEI 500 units
    https://www.bioz.com/result/restriction enzyme bpuei/product/New England Biolabs
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    restriction enzyme bpuei - by Bioz Stars, 2020-09
    93/100 stars

    Related Products / Commonly Used Together

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    Images

    1) Product Images from "FANCI and FANCD2 have common as well as independent functions during the cellular replication stress response"

    Article Title: FANCI and FANCD2 have common as well as independent functions during the cellular replication stress response

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkx847

    Confirmation of D2 −/− , I −/− and ID2 DKO cell lines. ( A ) PCR genotyping of D2 −/− and I −/− cells and targeting intermediates. Left panel : analyses of DNA fragments for WT (lane 1), D2 flox/Neo (lanes 2 and 3) and D2 −/− (lanes 4 and 5) after PCR amplification with primers FANCD2_EX11SF and FANCD2_LoxPSR flanking the targeted exon. Right panel : analyses of DNA fragments after PCR amplification from WT (lane 1), I flox/+ (lane 2) and I −/− (lanes 3 and 4) cells using primers FancIc_GG_LIF and FancIcond_GG_LoxR flanking the targeted exon. The PCR amplification spanning the targeted exons (exon 12 in FANCD2 ; exon 10 in FANCI ) was used to confirm the removal of the respective exon in the D2 −/− and I −/− cell lines. M: 1 kb markers. ( B ) Schematic of the FANCD2 targeting strategy in I −/− cells. CRISPR/Cas9-mediated gene targeting was used to functionally inactivate FANCD2 in the I −/− cell line. A guide RNA (purple sequence) was designed targeting FANCD2 exon 11 with the Cas9 cut site (red arrow) overlapping an endogenous BpuEI restriction enzyme recognition site (black bar). The PAM sequence of the sgRNA is shown in blue. Indels introduced at the Cas9 cut site should disrupt the BpuEI cleavage site. ( C ) Genotyping of ID2 DKO cells. PCR amplification and BpuEI restriction enzyme digestion of FANCD2 exon 11 in WT, I −/− and two ID2 −/− clones ( 1 and 2 ). Analyses of DNA fragments after PCR amplification with primers FancD2_CC_ F2 and FancD2_CC_ R2 (blue arrows from panel B) from WT (lanes 1 and 2), I −/− (lanes 3 and 4) and ID2 DKO cells (lanes 5, 6, 7 and 8) that had been untreated (lanes 1, 3, 5 and 7) or treated with BpuEI (lanes 2, 4, 6 and 8). Cleavage by BpuEI produces two faster migrating fragments (D, lanes 2 and 4). Resistance (R) to BpuEI digestion is seen in lanes 6 and 8 with the two ID2 DKO clones. ( D ) Sequence confirmation of CRISPR/Cas9 induced bi-allelic frameshift mutations in FANCD2 in the two ID2 DKO clones #1 and #2.
    Figure Legend Snippet: Confirmation of D2 −/− , I −/− and ID2 DKO cell lines. ( A ) PCR genotyping of D2 −/− and I −/− cells and targeting intermediates. Left panel : analyses of DNA fragments for WT (lane 1), D2 flox/Neo (lanes 2 and 3) and D2 −/− (lanes 4 and 5) after PCR amplification with primers FANCD2_EX11SF and FANCD2_LoxPSR flanking the targeted exon. Right panel : analyses of DNA fragments after PCR amplification from WT (lane 1), I flox/+ (lane 2) and I −/− (lanes 3 and 4) cells using primers FancIc_GG_LIF and FancIcond_GG_LoxR flanking the targeted exon. The PCR amplification spanning the targeted exons (exon 12 in FANCD2 ; exon 10 in FANCI ) was used to confirm the removal of the respective exon in the D2 −/− and I −/− cell lines. M: 1 kb markers. ( B ) Schematic of the FANCD2 targeting strategy in I −/− cells. CRISPR/Cas9-mediated gene targeting was used to functionally inactivate FANCD2 in the I −/− cell line. A guide RNA (purple sequence) was designed targeting FANCD2 exon 11 with the Cas9 cut site (red arrow) overlapping an endogenous BpuEI restriction enzyme recognition site (black bar). The PAM sequence of the sgRNA is shown in blue. Indels introduced at the Cas9 cut site should disrupt the BpuEI cleavage site. ( C ) Genotyping of ID2 DKO cells. PCR amplification and BpuEI restriction enzyme digestion of FANCD2 exon 11 in WT, I −/− and two ID2 −/− clones ( 1 and 2 ). Analyses of DNA fragments after PCR amplification with primers FancD2_CC_ F2 and FancD2_CC_ R2 (blue arrows from panel B) from WT (lanes 1 and 2), I −/− (lanes 3 and 4) and ID2 DKO cells (lanes 5, 6, 7 and 8) that had been untreated (lanes 1, 3, 5 and 7) or treated with BpuEI (lanes 2, 4, 6 and 8). Cleavage by BpuEI produces two faster migrating fragments (D, lanes 2 and 4). Resistance (R) to BpuEI digestion is seen in lanes 6 and 8 with the two ID2 DKO clones. ( D ) Sequence confirmation of CRISPR/Cas9 induced bi-allelic frameshift mutations in FANCD2 in the two ID2 DKO clones #1 and #2.

    Techniques Used: Polymerase Chain Reaction, Amplification, CRISPR, Sequencing, Clone Assay

    Related Articles

    Polymerase Chain Reaction:

    Article Title: FANCI and FANCD2 have common as well as independent functions during the cellular replication stress response
    Article Snippet: .. Two days after transfection, the cells were subcloned, and individual subclones were screened for targeting by PCR amplification of exon 11 and by subsequent digestion with the restriction enzyme BpuEI (New England BioLabs, Inc.). .. Clones that were resistant to digestion with BpuEI were TOPO TA cloned (Life Technologies).

    Transfection:

    Article Title: FANCI and FANCD2 have common as well as independent functions during the cellular replication stress response
    Article Snippet: .. Two days after transfection, the cells were subcloned, and individual subclones were screened for targeting by PCR amplification of exon 11 and by subsequent digestion with the restriction enzyme BpuEI (New England BioLabs, Inc.). .. Clones that were resistant to digestion with BpuEI were TOPO TA cloned (Life Technologies).

    Amplification:

    Article Title: FANCI and FANCD2 have common as well as independent functions during the cellular replication stress response
    Article Snippet: .. Two days after transfection, the cells were subcloned, and individual subclones were screened for targeting by PCR amplification of exon 11 and by subsequent digestion with the restriction enzyme BpuEI (New England BioLabs, Inc.). .. Clones that were resistant to digestion with BpuEI were TOPO TA cloned (Life Technologies).

    Purification:

    Article Title: Comprehensive Mutagenesis of Herpes Simplex Virus 1 Genome Identifies UL42 as an Inhibitor of Type I Interferon Induction
    Article Snippet: .. Purified viral genomes were digested with BpuEI (NEB), and annealed adapter oligonucleotides were ligated overnight at 16°C using T4 ligase (NEB). .. The sequencing library was constructed using a two-step Nextera indexing protocol with primers specific for the transposon and the adapter oligonucleotide (Illumina).

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    New England Biolabs restriction enzyme bpuei
    Confirmation of D2 −/− , I −/− and ID2 DKO cell lines. ( A ) <t>PCR</t> genotyping of D2 −/− and I −/− cells and targeting intermediates. Left panel : analyses of DNA fragments for WT (lane 1), D2 flox/Neo (lanes 2 and 3) and D2 −/− (lanes 4 and 5) after PCR amplification with primers FANCD2_EX11SF and FANCD2_LoxPSR flanking the targeted exon. Right panel : analyses of DNA fragments after PCR amplification from WT (lane 1), I flox/+ (lane 2) and I −/− (lanes 3 and 4) cells using primers FancIc_GG_LIF and FancIcond_GG_LoxR flanking the targeted exon. The PCR amplification spanning the targeted exons (exon 12 in FANCD2 ; exon 10 in FANCI ) was used to confirm the removal of the respective exon in the D2 −/− and I −/− cell lines. M: 1 kb markers. ( B ) Schematic of the FANCD2 targeting strategy in I −/− cells. CRISPR/Cas9-mediated gene targeting was used to functionally inactivate FANCD2 in the I −/− cell line. A guide RNA (purple sequence) was designed targeting FANCD2 exon 11 with the Cas9 cut site (red arrow) overlapping an endogenous <t>BpuEI</t> restriction enzyme recognition site (black bar). The PAM sequence of the sgRNA is shown in blue. Indels introduced at the Cas9 cut site should disrupt the BpuEI cleavage site. ( C ) Genotyping of ID2 DKO cells. PCR amplification and BpuEI restriction enzyme digestion of FANCD2 exon 11 in WT, I −/− and two ID2 −/− clones ( 1 and 2 ). Analyses of DNA fragments after PCR amplification with primers FancD2_CC_ F2 and FancD2_CC_ R2 (blue arrows from panel B) from WT (lanes 1 and 2), I −/− (lanes 3 and 4) and ID2 DKO cells (lanes 5, 6, 7 and 8) that had been untreated (lanes 1, 3, 5 and 7) or treated with BpuEI (lanes 2, 4, 6 and 8). Cleavage by BpuEI produces two faster migrating fragments (D, lanes 2 and 4). Resistance (R) to BpuEI digestion is seen in lanes 6 and 8 with the two ID2 DKO clones. ( D ) Sequence confirmation of CRISPR/Cas9 induced bi-allelic frameshift mutations in FANCD2 in the two ID2 DKO clones #1 and #2.
    Restriction Enzyme Bpuei, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/restriction enzyme bpuei/product/New England Biolabs
    Average 93 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    restriction enzyme bpuei - by Bioz Stars, 2020-09
    93/100 stars
      Buy from Supplier

    Image Search Results


    Confirmation of D2 −/− , I −/− and ID2 DKO cell lines. ( A ) PCR genotyping of D2 −/− and I −/− cells and targeting intermediates. Left panel : analyses of DNA fragments for WT (lane 1), D2 flox/Neo (lanes 2 and 3) and D2 −/− (lanes 4 and 5) after PCR amplification with primers FANCD2_EX11SF and FANCD2_LoxPSR flanking the targeted exon. Right panel : analyses of DNA fragments after PCR amplification from WT (lane 1), I flox/+ (lane 2) and I −/− (lanes 3 and 4) cells using primers FancIc_GG_LIF and FancIcond_GG_LoxR flanking the targeted exon. The PCR amplification spanning the targeted exons (exon 12 in FANCD2 ; exon 10 in FANCI ) was used to confirm the removal of the respective exon in the D2 −/− and I −/− cell lines. M: 1 kb markers. ( B ) Schematic of the FANCD2 targeting strategy in I −/− cells. CRISPR/Cas9-mediated gene targeting was used to functionally inactivate FANCD2 in the I −/− cell line. A guide RNA (purple sequence) was designed targeting FANCD2 exon 11 with the Cas9 cut site (red arrow) overlapping an endogenous BpuEI restriction enzyme recognition site (black bar). The PAM sequence of the sgRNA is shown in blue. Indels introduced at the Cas9 cut site should disrupt the BpuEI cleavage site. ( C ) Genotyping of ID2 DKO cells. PCR amplification and BpuEI restriction enzyme digestion of FANCD2 exon 11 in WT, I −/− and two ID2 −/− clones ( 1 and 2 ). Analyses of DNA fragments after PCR amplification with primers FancD2_CC_ F2 and FancD2_CC_ R2 (blue arrows from panel B) from WT (lanes 1 and 2), I −/− (lanes 3 and 4) and ID2 DKO cells (lanes 5, 6, 7 and 8) that had been untreated (lanes 1, 3, 5 and 7) or treated with BpuEI (lanes 2, 4, 6 and 8). Cleavage by BpuEI produces two faster migrating fragments (D, lanes 2 and 4). Resistance (R) to BpuEI digestion is seen in lanes 6 and 8 with the two ID2 DKO clones. ( D ) Sequence confirmation of CRISPR/Cas9 induced bi-allelic frameshift mutations in FANCD2 in the two ID2 DKO clones #1 and #2.

    Journal: Nucleic Acids Research

    Article Title: FANCI and FANCD2 have common as well as independent functions during the cellular replication stress response

    doi: 10.1093/nar/gkx847

    Figure Lengend Snippet: Confirmation of D2 −/− , I −/− and ID2 DKO cell lines. ( A ) PCR genotyping of D2 −/− and I −/− cells and targeting intermediates. Left panel : analyses of DNA fragments for WT (lane 1), D2 flox/Neo (lanes 2 and 3) and D2 −/− (lanes 4 and 5) after PCR amplification with primers FANCD2_EX11SF and FANCD2_LoxPSR flanking the targeted exon. Right panel : analyses of DNA fragments after PCR amplification from WT (lane 1), I flox/+ (lane 2) and I −/− (lanes 3 and 4) cells using primers FancIc_GG_LIF and FancIcond_GG_LoxR flanking the targeted exon. The PCR amplification spanning the targeted exons (exon 12 in FANCD2 ; exon 10 in FANCI ) was used to confirm the removal of the respective exon in the D2 −/− and I −/− cell lines. M: 1 kb markers. ( B ) Schematic of the FANCD2 targeting strategy in I −/− cells. CRISPR/Cas9-mediated gene targeting was used to functionally inactivate FANCD2 in the I −/− cell line. A guide RNA (purple sequence) was designed targeting FANCD2 exon 11 with the Cas9 cut site (red arrow) overlapping an endogenous BpuEI restriction enzyme recognition site (black bar). The PAM sequence of the sgRNA is shown in blue. Indels introduced at the Cas9 cut site should disrupt the BpuEI cleavage site. ( C ) Genotyping of ID2 DKO cells. PCR amplification and BpuEI restriction enzyme digestion of FANCD2 exon 11 in WT, I −/− and two ID2 −/− clones ( 1 and 2 ). Analyses of DNA fragments after PCR amplification with primers FancD2_CC_ F2 and FancD2_CC_ R2 (blue arrows from panel B) from WT (lanes 1 and 2), I −/− (lanes 3 and 4) and ID2 DKO cells (lanes 5, 6, 7 and 8) that had been untreated (lanes 1, 3, 5 and 7) or treated with BpuEI (lanes 2, 4, 6 and 8). Cleavage by BpuEI produces two faster migrating fragments (D, lanes 2 and 4). Resistance (R) to BpuEI digestion is seen in lanes 6 and 8 with the two ID2 DKO clones. ( D ) Sequence confirmation of CRISPR/Cas9 induced bi-allelic frameshift mutations in FANCD2 in the two ID2 DKO clones #1 and #2.

    Article Snippet: Two days after transfection, the cells were subcloned, and individual subclones were screened for targeting by PCR amplification of exon 11 and by subsequent digestion with the restriction enzyme BpuEI (New England BioLabs, Inc.).

    Techniques: Polymerase Chain Reaction, Amplification, CRISPR, Sequencing, Clone Assay