bsaxi  (New England Biolabs)


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    Name:
    BsaXI
    Description:
    BsaXI 500 units
    Catalog Number:
    r0609l
    Price:
    282
    Size:
    500 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs bsaxi
    BsaXI
    BsaXI 500 units
    https://www.bioz.com/result/bsaxi/product/New England Biolabs
    Average 96 stars, based on 5741 article reviews
    Price from $9.99 to $1999.99
    bsaxi - by Bioz Stars, 2020-09
    96/100 stars

    Images

    1) Product Images from "Genome editing to model and reverse a prevalent mutation associated with myeloproliferative neoplasms"

    Article Title: Genome editing to model and reverse a prevalent mutation associated with myeloproliferative neoplasms

    Journal: bioRxiv

    doi: 10.1101/855452

    Characterization of JAK2 V617F and ‘corrected’ F617V HUDEP clones. (a) DNA gel showing screening of V617F clones by PCR and BsaXI restriction digest. Lower band corresponds to WT allele and higher undigested fragment corresponds to V617F allele. (b) Sanger sequencing traces of V617F homozygous (F1) and heterozygous (H1) clones. (c) Viability curve depicting cell death in the absence of EPO. F1 and F3 V617F homozygote clones exhibited mildly higher cell viability than WT cells. Data is from n=5 independent biological replicates. Mean of all experiments ± SD shown. (d) . Sanger sequencing traces of F617V homozygous (cF1) and heterozygous (cH2) clones. (e) Immunoblot and signal intensity quantification show elevated phosphorylated STAT5 (P-STAT5) and uniform JAK2 expression in JAK2 V617F HUDEP-2 clones without erythropoietin (EPO). Signals were normalized to STAT5 and GAPDH. White bars, JAK2 WT clones; light pink bar, JAK2 V617F heterozygous clone; magenta bars, JAK2 V617F homozygous clones. (f) Immunoblot and signal intensity quantification show elevated phosphorylated STAT1 (P-STAT1) expression in JAK2 V617F HUDEP-2 clones both with and without erythropoietin (EPO). Signals were normalized to STAT1 and GAPDH. White bars, JAK2 WT clones; light pink bars, JAK2 V617F heterozygous clones; magenta bars, JAK2 V617F homozygous clones. (g) Representative FACS plots for gating live HUDEP-2s expressing Kusabira Orange, a marker gene indicative of viable HUDEP-2s. (h) Levels of Glycophorin A (GlyA), an erythroid-specific cell surface marker, in undifferentiated or fully differentiated HUDEP-2s at day 0 and day 5, respectively.
    Figure Legend Snippet: Characterization of JAK2 V617F and ‘corrected’ F617V HUDEP clones. (a) DNA gel showing screening of V617F clones by PCR and BsaXI restriction digest. Lower band corresponds to WT allele and higher undigested fragment corresponds to V617F allele. (b) Sanger sequencing traces of V617F homozygous (F1) and heterozygous (H1) clones. (c) Viability curve depicting cell death in the absence of EPO. F1 and F3 V617F homozygote clones exhibited mildly higher cell viability than WT cells. Data is from n=5 independent biological replicates. Mean of all experiments ± SD shown. (d) . Sanger sequencing traces of F617V homozygous (cF1) and heterozygous (cH2) clones. (e) Immunoblot and signal intensity quantification show elevated phosphorylated STAT5 (P-STAT5) and uniform JAK2 expression in JAK2 V617F HUDEP-2 clones without erythropoietin (EPO). Signals were normalized to STAT5 and GAPDH. White bars, JAK2 WT clones; light pink bar, JAK2 V617F heterozygous clone; magenta bars, JAK2 V617F homozygous clones. (f) Immunoblot and signal intensity quantification show elevated phosphorylated STAT1 (P-STAT1) expression in JAK2 V617F HUDEP-2 clones both with and without erythropoietin (EPO). Signals were normalized to STAT1 and GAPDH. White bars, JAK2 WT clones; light pink bars, JAK2 V617F heterozygous clones; magenta bars, JAK2 V617F homozygous clones. (g) Representative FACS plots for gating live HUDEP-2s expressing Kusabira Orange, a marker gene indicative of viable HUDEP-2s. (h) Levels of Glycophorin A (GlyA), an erythroid-specific cell surface marker, in undifferentiated or fully differentiated HUDEP-2s at day 0 and day 5, respectively.

    Techniques Used: Clone Assay, Polymerase Chain Reaction, Sequencing, Expressing, FACS, Marker

    2) Product Images from "DNA polymerase ? and PARP activities in base excision repair in living cells"

    Article Title: DNA polymerase ? and PARP activities in base excision repair in living cells

    Journal: DNA repair

    doi: 10.1016/j.dnarep.2009.08.004

    (A) Transcriptional base substitution in the luciferase gene for the study of SN-BER. The codon 27 of the luciferase gene was modified to a stop codon by introduction of a uracil residue, as indicated. The uracil will be converted to a leucine codon after DNA repair, resulting in luciferase protein expression. (B) The construction of plasmid DNA containing uracil is described under “Materials and methods.” The Renilla luciferase gene of pGL4.75 was replaced by the Chroma-Luc™ gene, and the oligonucleotide fragment containing uracil was ligated at the Bsa XI site introduced by site-directed mutagenesis in the Chroma-Luc™ gene. (C) Confirmation of plasmid preparations. Plasmids AM1, AM1-P (positive) and AM1-U (uracil) were treated with Bsa XI at 37°C for 1 h, and then the mixtures were analyzed by 1% agarose gel electrophoresis. AM1 was used as reference for closed circular (lane 1) and linear DNA (lane 2).
    Figure Legend Snippet: (A) Transcriptional base substitution in the luciferase gene for the study of SN-BER. The codon 27 of the luciferase gene was modified to a stop codon by introduction of a uracil residue, as indicated. The uracil will be converted to a leucine codon after DNA repair, resulting in luciferase protein expression. (B) The construction of plasmid DNA containing uracil is described under “Materials and methods.” The Renilla luciferase gene of pGL4.75 was replaced by the Chroma-Luc™ gene, and the oligonucleotide fragment containing uracil was ligated at the Bsa XI site introduced by site-directed mutagenesis in the Chroma-Luc™ gene. (C) Confirmation of plasmid preparations. Plasmids AM1, AM1-P (positive) and AM1-U (uracil) were treated with Bsa XI at 37°C for 1 h, and then the mixtures were analyzed by 1% agarose gel electrophoresis. AM1 was used as reference for closed circular (lane 1) and linear DNA (lane 2).

    Techniques Used: Luciferase, Modification, Expressing, Plasmid Preparation, Mutagenesis, Agarose Gel Electrophoresis

    3) Product Images from "Screening for hotspot mutations in PI3K, JAK2, FLT3 and NPM1 in patients with myelodysplastic syndromes"

    Article Title: Screening for hotspot mutations in PI3K, JAK2, FLT3 and NPM1 in patients with myelodysplastic syndromes

    Journal: Clinics

    doi: 10.1590/S1807-59322011000500014

    JAK2 V617F genotyping. (A) PCR amplification of JAK2 : lane 1: 100 bp ladder; lane 2: negative control; lanes 3 to 6 – 460-bp amplicons obtained from the genomic DNA of a patient with PV (3), a CMML patient after disease progression (4) and two MDS patients (with RA) (5 and 6). (B) BsaXI digestion: lane 1: 100 bp ladder, lane 2: negative control; lanes 3 and 4: digestion pattern observed in a PV patient (3) and in the CMML patient positive for the JAK2 V617F allele after disease progression (4); lanes 5 and 6: digestion pattern observed in two MDS patients (with RA) with wild-type JAK2 alleles.
    Figure Legend Snippet: JAK2 V617F genotyping. (A) PCR amplification of JAK2 : lane 1: 100 bp ladder; lane 2: negative control; lanes 3 to 6 – 460-bp amplicons obtained from the genomic DNA of a patient with PV (3), a CMML patient after disease progression (4) and two MDS patients (with RA) (5 and 6). (B) BsaXI digestion: lane 1: 100 bp ladder, lane 2: negative control; lanes 3 and 4: digestion pattern observed in a PV patient (3) and in the CMML patient positive for the JAK2 V617F allele after disease progression (4); lanes 5 and 6: digestion pattern observed in two MDS patients (with RA) with wild-type JAK2 alleles.

    Techniques Used: Polymerase Chain Reaction, Amplification, Negative Control

    Related Articles

    Amplification:

    Article Title: The haplotype of three polymorphisms in the SATB1 promoter region impacts survival in breast cancer patients
    Article Snippet: .. Following denaturation at 95°C, 38 cycles of DNA amplification were performed using Taq PCR Mastermix (Eppendorf, Hamburg, Germany) at 95°C for 40 sec, 62°C for 40 sec and 72°C for 40 sec. Digestion with Bsa XI (New England Biolabs, Inc., Ipswich, MA, USA) at 37°C resulted in fragments of 50, 30 and 27 bp for the C-allele versus 107 bp for the TT-genotype (no digestion). .. Electrophoresis was performed in 2.8% agarose gels using SYBR Safe® DNA gel stain (Invitrogen Life Technologies, Carlsbad, CA, USA) for visualization under ultraviolet light.

    Agarose Gel Electrophoresis:

    Article Title: DNA polymerase ? and PARP activities in base excision repair in living cells
    Article Snippet: .. The Bsa XI site was introduced in the Chroma-Luc™ gene by site-directed mutagenesis, and the resultant plasmid was digested by Bsa XI (New England Biolabs, Ipswich, MA) and purified with a gel extraction kit (Qiagen, Valencia, CA) after 1% agarose gel electrophoresis. .. In the descriptions to follow, the underlined residues illustrate the altered codon base pairs in the luciferase gene.

    Article Title: Screening for hotspot mutations in PI3K, JAK2, FLT3 and NPM1 in patients with myelodysplastic syndromes
    Article Snippet: .. For RFLP analysis, JAK2 and FLT3 PCR products were digested with BsaXI or Eco321 (New England Biolabs, Hitchin, UK), respectively, according to the manufacturer's protocol, and visualized on a 2.5% agarose gel. .. The normal genotype for JAK2 was represented by a 460-bp fragment, and the heterozygous genotype was represented by 460-bp, 241-bp and 189-bp fragments, whereas the homozygous mutant genotype produced 241-bp and 189-bp fragments.

    Article Title: Genome editing to model and reverse a prevalent mutation associated with myeloproliferative neoplasms
    Article Snippet: .. The PCR amplicons were subjected to restriction digestion using BsaXI (New England Biolabs) for 60 minutes at 37°C, and the digested products were run on a 1.5% agarose gel to visualize either the appearance (for WT and F617V clones) or disappearance (for V617F clones) of the digested products. .. Clones selected from the restriction digest screen were further verified by Sanger sequencing.

    Mutagenesis:

    Article Title: DNA polymerase ? and PARP activities in base excision repair in living cells
    Article Snippet: .. The Bsa XI site was introduced in the Chroma-Luc™ gene by site-directed mutagenesis, and the resultant plasmid was digested by Bsa XI (New England Biolabs, Ipswich, MA) and purified with a gel extraction kit (Qiagen, Valencia, CA) after 1% agarose gel electrophoresis. .. In the descriptions to follow, the underlined residues illustrate the altered codon base pairs in the luciferase gene.

    Size-exclusion Chromatography:

    Article Title: The haplotype of three polymorphisms in the SATB1 promoter region impacts survival in breast cancer patients
    Article Snippet: .. Following denaturation at 95°C, 38 cycles of DNA amplification were performed using Taq PCR Mastermix (Eppendorf, Hamburg, Germany) at 95°C for 40 sec, 62°C for 40 sec and 72°C for 40 sec. Digestion with Bsa XI (New England Biolabs, Inc., Ipswich, MA, USA) at 37°C resulted in fragments of 50, 30 and 27 bp for the C-allele versus 107 bp for the TT-genotype (no digestion). .. Electrophoresis was performed in 2.8% agarose gels using SYBR Safe® DNA gel stain (Invitrogen Life Technologies, Carlsbad, CA, USA) for visualization under ultraviolet light.

    Purification:

    Article Title: DNA polymerase ? and PARP activities in base excision repair in living cells
    Article Snippet: .. The Bsa XI site was introduced in the Chroma-Luc™ gene by site-directed mutagenesis, and the resultant plasmid was digested by Bsa XI (New England Biolabs, Ipswich, MA) and purified with a gel extraction kit (Qiagen, Valencia, CA) after 1% agarose gel electrophoresis. .. In the descriptions to follow, the underlined residues illustrate the altered codon base pairs in the luciferase gene.

    Polymerase Chain Reaction:

    Article Title: Advantages of the quenching probe method over other PCR-based methods for detection of the JAK2 V617F mutation
    Article Snippet: .. The resulting products were digested with Bsa XI (New England Biolabs, Hitchin, UK), and the PCR products and Bsa XI-treated products were electrophoresed as described above. .. HRM was performed using a LightCycler Nano (Roche Diagnostics, Mannheim, Germany) according to a previously reported protocol, with modifications ( ).

    Article Title: Mitochondrial DNA Mutations Induced by Carbon Ions Radiation: A Preliminary Study
    Article Snippet: .. The PCR product of 5 µL was mixed with 3 U of BsaXI (New England Biolabs, Frankfurt, Germany) in 1× buffer containing 50 mmol/L potassium acetate, 20 mmol/L Tris–acetate, 10 mmol/L magnesium acetate, 1 mmol/L Dithiothreitol (DTT), and pH 7.9. ..

    Article Title: The haplotype of three polymorphisms in the SATB1 promoter region impacts survival in breast cancer patients
    Article Snippet: .. Following denaturation at 95°C, 38 cycles of DNA amplification were performed using Taq PCR Mastermix (Eppendorf, Hamburg, Germany) at 95°C for 40 sec, 62°C for 40 sec and 72°C for 40 sec. Digestion with Bsa XI (New England Biolabs, Inc., Ipswich, MA, USA) at 37°C resulted in fragments of 50, 30 and 27 bp for the C-allele versus 107 bp for the TT-genotype (no digestion). .. Electrophoresis was performed in 2.8% agarose gels using SYBR Safe® DNA gel stain (Invitrogen Life Technologies, Carlsbad, CA, USA) for visualization under ultraviolet light.

    Article Title: Screening for hotspot mutations in PI3K, JAK2, FLT3 and NPM1 in patients with myelodysplastic syndromes
    Article Snippet: .. For RFLP analysis, JAK2 and FLT3 PCR products were digested with BsaXI or Eco321 (New England Biolabs, Hitchin, UK), respectively, according to the manufacturer's protocol, and visualized on a 2.5% agarose gel. .. The normal genotype for JAK2 was represented by a 460-bp fragment, and the heterozygous genotype was represented by 460-bp, 241-bp and 189-bp fragments, whereas the homozygous mutant genotype produced 241-bp and 189-bp fragments.

    Article Title: Genome editing to model and reverse a prevalent mutation associated with myeloproliferative neoplasms
    Article Snippet: .. The PCR amplicons were subjected to restriction digestion using BsaXI (New England Biolabs) for 60 minutes at 37°C, and the digested products were run on a 1.5% agarose gel to visualize either the appearance (for WT and F617V clones) or disappearance (for V617F clones) of the digested products. .. Clones selected from the restriction digest screen were further verified by Sanger sequencing.

    Gel Extraction:

    Article Title: DNA polymerase ? and PARP activities in base excision repair in living cells
    Article Snippet: .. The Bsa XI site was introduced in the Chroma-Luc™ gene by site-directed mutagenesis, and the resultant plasmid was digested by Bsa XI (New England Biolabs, Ipswich, MA) and purified with a gel extraction kit (Qiagen, Valencia, CA) after 1% agarose gel electrophoresis. .. In the descriptions to follow, the underlined residues illustrate the altered codon base pairs in the luciferase gene.

    Plasmid Preparation:

    Article Title: DNA polymerase ? and PARP activities in base excision repair in living cells
    Article Snippet: .. The Bsa XI site was introduced in the Chroma-Luc™ gene by site-directed mutagenesis, and the resultant plasmid was digested by Bsa XI (New England Biolabs, Ipswich, MA) and purified with a gel extraction kit (Qiagen, Valencia, CA) after 1% agarose gel electrophoresis. .. In the descriptions to follow, the underlined residues illustrate the altered codon base pairs in the luciferase gene.

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  • 96
    New England Biolabs bsaxi
    Characterization of JAK2 V617F and ‘corrected’ F617V HUDEP clones. (a) DNA gel showing screening of V617F clones by <t>PCR</t> and <t>BsaXI</t> restriction digest. Lower band corresponds to WT allele and higher undigested fragment corresponds to V617F allele. (b) Sanger sequencing traces of V617F homozygous (F1) and heterozygous (H1) clones. (c) Viability curve depicting cell death in the absence of EPO. F1 and F3 V617F homozygote clones exhibited mildly higher cell viability than WT cells. Data is from n=5 independent biological replicates. Mean of all experiments ± SD shown. (d) . Sanger sequencing traces of F617V homozygous (cF1) and heterozygous (cH2) clones. (e) Immunoblot and signal intensity quantification show elevated phosphorylated STAT5 (P-STAT5) and uniform JAK2 expression in JAK2 V617F HUDEP-2 clones without erythropoietin (EPO). Signals were normalized to STAT5 and GAPDH. White bars, JAK2 WT clones; light pink bar, JAK2 V617F heterozygous clone; magenta bars, JAK2 V617F homozygous clones. (f) Immunoblot and signal intensity quantification show elevated phosphorylated STAT1 (P-STAT1) expression in JAK2 V617F HUDEP-2 clones both with and without erythropoietin (EPO). Signals were normalized to STAT1 and GAPDH. White bars, JAK2 WT clones; light pink bars, JAK2 V617F heterozygous clones; magenta bars, JAK2 V617F homozygous clones. (g) Representative FACS plots for gating live HUDEP-2s expressing Kusabira Orange, a marker gene indicative of viable HUDEP-2s. (h) Levels of Glycophorin A (GlyA), an erythroid-specific cell surface marker, in undifferentiated or fully differentiated HUDEP-2s at day 0 and day 5, respectively.
    Bsaxi, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsaxi/product/New England Biolabs
    Average 96 stars, based on 31 article reviews
    Price from $9.99 to $1999.99
    bsaxi - by Bioz Stars, 2020-09
    96/100 stars
      Buy from Supplier

    Image Search Results


    Characterization of JAK2 V617F and ‘corrected’ F617V HUDEP clones. (a) DNA gel showing screening of V617F clones by PCR and BsaXI restriction digest. Lower band corresponds to WT allele and higher undigested fragment corresponds to V617F allele. (b) Sanger sequencing traces of V617F homozygous (F1) and heterozygous (H1) clones. (c) Viability curve depicting cell death in the absence of EPO. F1 and F3 V617F homozygote clones exhibited mildly higher cell viability than WT cells. Data is from n=5 independent biological replicates. Mean of all experiments ± SD shown. (d) . Sanger sequencing traces of F617V homozygous (cF1) and heterozygous (cH2) clones. (e) Immunoblot and signal intensity quantification show elevated phosphorylated STAT5 (P-STAT5) and uniform JAK2 expression in JAK2 V617F HUDEP-2 clones without erythropoietin (EPO). Signals were normalized to STAT5 and GAPDH. White bars, JAK2 WT clones; light pink bar, JAK2 V617F heterozygous clone; magenta bars, JAK2 V617F homozygous clones. (f) Immunoblot and signal intensity quantification show elevated phosphorylated STAT1 (P-STAT1) expression in JAK2 V617F HUDEP-2 clones both with and without erythropoietin (EPO). Signals were normalized to STAT1 and GAPDH. White bars, JAK2 WT clones; light pink bars, JAK2 V617F heterozygous clones; magenta bars, JAK2 V617F homozygous clones. (g) Representative FACS plots for gating live HUDEP-2s expressing Kusabira Orange, a marker gene indicative of viable HUDEP-2s. (h) Levels of Glycophorin A (GlyA), an erythroid-specific cell surface marker, in undifferentiated or fully differentiated HUDEP-2s at day 0 and day 5, respectively.

    Journal: bioRxiv

    Article Title: Genome editing to model and reverse a prevalent mutation associated with myeloproliferative neoplasms

    doi: 10.1101/855452

    Figure Lengend Snippet: Characterization of JAK2 V617F and ‘corrected’ F617V HUDEP clones. (a) DNA gel showing screening of V617F clones by PCR and BsaXI restriction digest. Lower band corresponds to WT allele and higher undigested fragment corresponds to V617F allele. (b) Sanger sequencing traces of V617F homozygous (F1) and heterozygous (H1) clones. (c) Viability curve depicting cell death in the absence of EPO. F1 and F3 V617F homozygote clones exhibited mildly higher cell viability than WT cells. Data is from n=5 independent biological replicates. Mean of all experiments ± SD shown. (d) . Sanger sequencing traces of F617V homozygous (cF1) and heterozygous (cH2) clones. (e) Immunoblot and signal intensity quantification show elevated phosphorylated STAT5 (P-STAT5) and uniform JAK2 expression in JAK2 V617F HUDEP-2 clones without erythropoietin (EPO). Signals were normalized to STAT5 and GAPDH. White bars, JAK2 WT clones; light pink bar, JAK2 V617F heterozygous clone; magenta bars, JAK2 V617F homozygous clones. (f) Immunoblot and signal intensity quantification show elevated phosphorylated STAT1 (P-STAT1) expression in JAK2 V617F HUDEP-2 clones both with and without erythropoietin (EPO). Signals were normalized to STAT1 and GAPDH. White bars, JAK2 WT clones; light pink bars, JAK2 V617F heterozygous clones; magenta bars, JAK2 V617F homozygous clones. (g) Representative FACS plots for gating live HUDEP-2s expressing Kusabira Orange, a marker gene indicative of viable HUDEP-2s. (h) Levels of Glycophorin A (GlyA), an erythroid-specific cell surface marker, in undifferentiated or fully differentiated HUDEP-2s at day 0 and day 5, respectively.

    Article Snippet: The PCR amplicons were subjected to restriction digestion using BsaXI (New England Biolabs) for 60 minutes at 37°C, and the digested products were run on a 1.5% agarose gel to visualize either the appearance (for WT and F617V clones) or disappearance (for V617F clones) of the digested products.

    Techniques: Clone Assay, Polymerase Chain Reaction, Sequencing, Expressing, FACS, Marker

    (A) Transcriptional base substitution in the luciferase gene for the study of SN-BER. The codon 27 of the luciferase gene was modified to a stop codon by introduction of a uracil residue, as indicated. The uracil will be converted to a leucine codon after DNA repair, resulting in luciferase protein expression. (B) The construction of plasmid DNA containing uracil is described under “Materials and methods.” The Renilla luciferase gene of pGL4.75 was replaced by the Chroma-Luc™ gene, and the oligonucleotide fragment containing uracil was ligated at the Bsa XI site introduced by site-directed mutagenesis in the Chroma-Luc™ gene. (C) Confirmation of plasmid preparations. Plasmids AM1, AM1-P (positive) and AM1-U (uracil) were treated with Bsa XI at 37°C for 1 h, and then the mixtures were analyzed by 1% agarose gel electrophoresis. AM1 was used as reference for closed circular (lane 1) and linear DNA (lane 2).

    Journal: DNA repair

    Article Title: DNA polymerase ? and PARP activities in base excision repair in living cells

    doi: 10.1016/j.dnarep.2009.08.004

    Figure Lengend Snippet: (A) Transcriptional base substitution in the luciferase gene for the study of SN-BER. The codon 27 of the luciferase gene was modified to a stop codon by introduction of a uracil residue, as indicated. The uracil will be converted to a leucine codon after DNA repair, resulting in luciferase protein expression. (B) The construction of plasmid DNA containing uracil is described under “Materials and methods.” The Renilla luciferase gene of pGL4.75 was replaced by the Chroma-Luc™ gene, and the oligonucleotide fragment containing uracil was ligated at the Bsa XI site introduced by site-directed mutagenesis in the Chroma-Luc™ gene. (C) Confirmation of plasmid preparations. Plasmids AM1, AM1-P (positive) and AM1-U (uracil) were treated with Bsa XI at 37°C for 1 h, and then the mixtures were analyzed by 1% agarose gel electrophoresis. AM1 was used as reference for closed circular (lane 1) and linear DNA (lane 2).

    Article Snippet: The Bsa XI site was introduced in the Chroma-Luc™ gene by site-directed mutagenesis, and the resultant plasmid was digested by Bsa XI (New England Biolabs, Ipswich, MA) and purified with a gel extraction kit (Qiagen, Valencia, CA) after 1% agarose gel electrophoresis.

    Techniques: Luciferase, Modification, Expressing, Plasmid Preparation, Mutagenesis, Agarose Gel Electrophoresis

    JAK2 V617F genotyping. (A) PCR amplification of JAK2 : lane 1: 100 bp ladder; lane 2: negative control; lanes 3 to 6 – 460-bp amplicons obtained from the genomic DNA of a patient with PV (3), a CMML patient after disease progression (4) and two MDS patients (with RA) (5 and 6). (B) BsaXI digestion: lane 1: 100 bp ladder, lane 2: negative control; lanes 3 and 4: digestion pattern observed in a PV patient (3) and in the CMML patient positive for the JAK2 V617F allele after disease progression (4); lanes 5 and 6: digestion pattern observed in two MDS patients (with RA) with wild-type JAK2 alleles.

    Journal: Clinics

    Article Title: Screening for hotspot mutations in PI3K, JAK2, FLT3 and NPM1 in patients with myelodysplastic syndromes

    doi: 10.1590/S1807-59322011000500014

    Figure Lengend Snippet: JAK2 V617F genotyping. (A) PCR amplification of JAK2 : lane 1: 100 bp ladder; lane 2: negative control; lanes 3 to 6 – 460-bp amplicons obtained from the genomic DNA of a patient with PV (3), a CMML patient after disease progression (4) and two MDS patients (with RA) (5 and 6). (B) BsaXI digestion: lane 1: 100 bp ladder, lane 2: negative control; lanes 3 and 4: digestion pattern observed in a PV patient (3) and in the CMML patient positive for the JAK2 V617F allele after disease progression (4); lanes 5 and 6: digestion pattern observed in two MDS patients (with RA) with wild-type JAK2 alleles.

    Article Snippet: For RFLP analysis, JAK2 and FLT3 PCR products were digested with BsaXI or Eco321 (New England Biolabs, Hitchin, UK), respectively, according to the manufacturer's protocol, and visualized on a 2.5% agarose gel.

    Techniques: Polymerase Chain Reaction, Amplification, Negative Control