nuclease bal 31  (New England Biolabs)


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  • 99
    Name:
    Nuclease BAL 31
    Description:
    Nuclease BAL 31 50 units
    Catalog Number:
    m0213s
    Price:
    68
    Size:
    50 units
    Category:
    Exonucleases
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    Structured Review

    New England Biolabs nuclease bal 31
    Nuclease BAL 31
    Nuclease BAL 31 50 units
    https://www.bioz.com/result/nuclease bal 31/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nuclease bal 31 - by Bioz Stars, 2020-04
    99/100 stars

    Images

    1) Product Images from "Structural diversity of supercoiled DNA"

    Article Title: Structural diversity of supercoiled DNA

    Journal: Nature Communications

    doi: 10.1038/ncomms9440

    Mapping Bal-31 cleavage. To determine whether Bal-31 cleavage occurs at multiple sites or at a preferred site, the Δ Lk =−6 topoisomer was cleaved with Bal-31 and various restriction enzymes. ( a ) Products were separated by agarose gel electrophoresis. Left, (lanes 1–5), control reactions, mc336 (approximately equal mixture of Δ Lk =−2 and Δ Lk =−3 topoisomers) with combinations of the various restriction enzymes (as indicated) to generate fragments of known DNA lengths. Right, (lanes 6–9), Δ Lk =−6 topoisomer cleaved first with Bal-31, followed by a restriction enzyme (as indicated). Mr 1 : 100 bp DNA ladder, Mr 2 : Low molecular weight DNA ladder. ( b ) Map of the minicircle sequence showing the positions of the restriction enzymes used, the estimated location of Bal-31 cleavage (with parentheses indicating the range), and the location of the observed base-pair breaking in MD simulation of the Δ Lk =−3 topoisomer.
    Figure Legend Snippet: Mapping Bal-31 cleavage. To determine whether Bal-31 cleavage occurs at multiple sites or at a preferred site, the Δ Lk =−6 topoisomer was cleaved with Bal-31 and various restriction enzymes. ( a ) Products were separated by agarose gel electrophoresis. Left, (lanes 1–5), control reactions, mc336 (approximately equal mixture of Δ Lk =−2 and Δ Lk =−3 topoisomers) with combinations of the various restriction enzymes (as indicated) to generate fragments of known DNA lengths. Right, (lanes 6–9), Δ Lk =−6 topoisomer cleaved first with Bal-31, followed by a restriction enzyme (as indicated). Mr 1 : 100 bp DNA ladder, Mr 2 : Low molecular weight DNA ladder. ( b ) Map of the minicircle sequence showing the positions of the restriction enzymes used, the estimated location of Bal-31 cleavage (with parentheses indicating the range), and the location of the observed base-pair breaking in MD simulation of the Δ Lk =−3 topoisomer.

    Techniques Used: Agarose Gel Electrophoresis, Molecular Weight, Sequencing

    Effect of supercoiling on DNA base accessibility. ( a ) Minicircle DNA incubated with nuclease Bal-31. Over time, samples were removed, quenched by the addition of stop buffer and the products analysed by polyacrylamide gel electrophoresis. Mr: 100 bp DNA ladder, L: linearized 336 bp DNA, N: nicked 336 bp minicircle. ( b ) Graphic representation of the data shown in ( a ) Fitted lines are for visualization purposes only. ( c ) MD simulation of the Δ Lk =−3 topoisomer in explicit solvent. Splayed bases were found at a sharp bend of a needle conformation. This may be a potential atomistic explanation for Bal-31 susceptibility of negatively supercoiled topoisomers.
    Figure Legend Snippet: Effect of supercoiling on DNA base accessibility. ( a ) Minicircle DNA incubated with nuclease Bal-31. Over time, samples were removed, quenched by the addition of stop buffer and the products analysed by polyacrylamide gel electrophoresis. Mr: 100 bp DNA ladder, L: linearized 336 bp DNA, N: nicked 336 bp minicircle. ( b ) Graphic representation of the data shown in ( a ) Fitted lines are for visualization purposes only. ( c ) MD simulation of the Δ Lk =−3 topoisomer in explicit solvent. Splayed bases were found at a sharp bend of a needle conformation. This may be a potential atomistic explanation for Bal-31 susceptibility of negatively supercoiled topoisomers.

    Techniques Used: Incubation, Polyacrylamide Gel Electrophoresis

    2) Product Images from "A shotgun antisense approach to the identification of novel essential genes in Pseudomonas aeruginosa"

    Article Title: A shotgun antisense approach to the identification of novel essential genes in Pseudomonas aeruginosa

    Journal: BMC Microbiology

    doi: 10.1186/1471-2180-14-24

    Construction and screening of PAO1 SALs. (A) Genomic DNA was isolated from P. aeruginosa PAO1 and nebulized to obtain sheared fragments of 200–800 bp. After treatment with exonuclease BAL-31 and Klenow polymerase, the genomic DNA fragments were cloned into the E. coli strain JM109, downstream of the arabinose-inducible promoter P BAD of the pHERD20T vector. (B) E. coli transformants, representing the PAO1 shotgun antisense library (SAL), were arrayed in 96-well microplates and (C) mated with P. aeruginosa PAO1 in the presence of a helper strain (triparental mating). (D) SAL recipient PAO1 exconjugants were selected by spotting on PIA plates supplemented with Cb both in the absence and in the presence of the P BAD inducer arabinose. Recipient PAO1 exconjugant spots were inspected for growth defects following 24 h of incubation at 37°C. (E) The identity of the genomic fragments eliciting growth defects (lethal effects, indicated by a lack of a spot: only with inducer, e.g. clones A4, A8, B5, and E4, and with and without an inducer, e.g. clones A2 and E6; growth impairment, indicated as gray spots: only with an inducer, e.g. clones C2, A6, and B6, and with and without an inducer, e.g. C3 and B8) was determined by sequencing the inserts in the corresponding clones of E. coli SAL.
    Figure Legend Snippet: Construction and screening of PAO1 SALs. (A) Genomic DNA was isolated from P. aeruginosa PAO1 and nebulized to obtain sheared fragments of 200–800 bp. After treatment with exonuclease BAL-31 and Klenow polymerase, the genomic DNA fragments were cloned into the E. coli strain JM109, downstream of the arabinose-inducible promoter P BAD of the pHERD20T vector. (B) E. coli transformants, representing the PAO1 shotgun antisense library (SAL), were arrayed in 96-well microplates and (C) mated with P. aeruginosa PAO1 in the presence of a helper strain (triparental mating). (D) SAL recipient PAO1 exconjugants were selected by spotting on PIA plates supplemented with Cb both in the absence and in the presence of the P BAD inducer arabinose. Recipient PAO1 exconjugant spots were inspected for growth defects following 24 h of incubation at 37°C. (E) The identity of the genomic fragments eliciting growth defects (lethal effects, indicated by a lack of a spot: only with inducer, e.g. clones A4, A8, B5, and E4, and with and without an inducer, e.g. clones A2 and E6; growth impairment, indicated as gray spots: only with an inducer, e.g. clones C2, A6, and B6, and with and without an inducer, e.g. C3 and B8) was determined by sequencing the inserts in the corresponding clones of E. coli SAL.

    Techniques Used: Isolation, Clone Assay, Plasmid Preparation, Incubation, Sequencing

    Related Articles

    DNA Extraction:

    Article Title: Brochothrix thermosphacta Bacteriophages Feature Heterogeneous and Highly Mosaic Genomes and Utilize Unique Prophage Insertion Sites ▿ Bacteriophages Feature Heterogeneous and Highly Mosaic Genomes and Utilize Unique Prophage Insertion Sites ▿ †
    Article Snippet: Paragraph title: Phage propagation and DNA isolation and manipulation. ... Bal31 nuclease treatment was performed by digestion of 40 μg of phage DNA with 20 U of Bal31 (New England Biolabs) at 30°C.

    Clone Assay:

    Article Title: The mitochondrial genome of the pathogenic yeast Candida subhashii: GC-rich linear DNA with a protein covalently attached to the 5? termini
    Article Snippet: Paragraph title: Cloning and analysis of the terminal mtDNA fragments. ... Approximately 1 μg mtDNA was treated with either exonuclease III (New England Biolabs) or BAL-31 nuclease (New England Biolabs) according to the manufacturer's instructions.

    Agarose Gel Electrophoresis:

    Article Title: The genome and proteome of Serratia bacteriophage ? which forms unstable lysogens
    Article Snippet: .. DNA sequencing and annotation Phage DNA was digested, ligated and treated with Bal31 exonuclease according to manufacturer’s specifications (NEB) and resolved using a 1% agarose gel. ..

    Article Title: The mitochondrial genome of the pathogenic yeast Candida subhashii: GC-rich linear DNA with a protein covalently attached to the 5? termini
    Article Snippet: Approximately 1 μg mtDNA was treated with either exonuclease III (New England Biolabs) or BAL-31 nuclease (New England Biolabs) according to the manufacturer's instructions. .. After heat inactivation of the enzyme, the mtDNA was extracted, digested with the restriction endonuclease Xba I (Takara) and electrophoretically separated on a 0.9 % agarose gel.

    Isolation:

    Article Title: The mitochondrial genome of the pathogenic yeast Candida subhashii: GC-rich linear DNA with a protein covalently attached to the 5? termini
    Article Snippet: Briefly, isolated mtDNA was treated with calf intestinal phosphatase (CIP) for 60 min at 37 °C. .. Approximately 1 μg mtDNA was treated with either exonuclease III (New England Biolabs) or BAL-31 nuclease (New England Biolabs) according to the manufacturer's instructions.

    Pulsed-Field Gel:

    Article Title: The genome and proteome of Serratia bacteriophage ? which forms unstable lysogens
    Article Snippet: DNA sequencing and annotation Phage DNA was digested, ligated and treated with Bal31 exonuclease according to manufacturer’s specifications (NEB) and resolved using a 1% agarose gel. .. Pulsed-field gel analysis was carried out following the protocol of Lingohr et al ., [ ].

    Article Title: Brochothrix thermosphacta Bacteriophages Feature Heterogeneous and Highly Mosaic Genomes and Utilize Unique Prophage Insertion Sites ▿ Bacteriophages Feature Heterogeneous and Highly Mosaic Genomes and Utilize Unique Prophage Insertion Sites ▿ †
    Article Snippet: Bal31 nuclease treatment was performed by digestion of 40 μg of phage DNA with 20 U of Bal31 (New England Biolabs) at 30°C. .. Pulsed-field gel electrophoresis (PFGE) was performed in 0.5× Tris-borate-EDTA buffer with an initial switch time of 1 s and a final switch time of 6 s at 5 V/cm, at an angle of 120° and a 14°C buffer temperature (CHEF DR III apparatus; Bio-Rad, Rainach, Switzerland) for 20 h. Lambda Mix 19 (Fermentas) and MidRange I PFG and MidRange II PFG markers (New England Biolabs) were used as DNA size standards.

    Incubation:

    Article Title: The mitochondrial genome of the pathogenic yeast Candida subhashii: GC-rich linear DNA with a protein covalently attached to the 5? termini
    Article Snippet: The enzyme was inhibited by incubation in the presence of 5 mM EDTA (pH 7.5) at 70 °C for 10 min, and mtDNA was then purified using a NucleoSpin Extract II column (Macherey-Nagel). .. Approximately 1 μg mtDNA was treated with either exonuclease III (New England Biolabs) or BAL-31 nuclease (New England Biolabs) according to the manufacturer's instructions.

    Article Title: Novel Virulent and Broad-Host-Range Erwinia amylovora Bacteriophages Reveal a High Degree of Mosaicism and a Relationship to Enterobacteriaceae Phages ▿ Phages ▿ †
    Article Snippet: .. Briefly, phage DNA was treated with BAL31 nuclease (NEB, Ipswich, MA) at 30°C for different incubation times prior to digestions with selected restriction enzymes. .. For the identification of potential cohesive ends ( cos ), half of the restriction-digested phage DNA was incubated at 62°C for 10 min to separate potentially ligated cos sites and immediately stored on ice.

    Purification:

    Article Title: The mitochondrial genome of the pathogenic yeast Candida subhashii: GC-rich linear DNA with a protein covalently attached to the 5? termini
    Article Snippet: The enzyme was inhibited by incubation in the presence of 5 mM EDTA (pH 7.5) at 70 °C for 10 min, and mtDNA was then purified using a NucleoSpin Extract II column (Macherey-Nagel). .. Approximately 1 μg mtDNA was treated with either exonuclease III (New England Biolabs) or BAL-31 nuclease (New England Biolabs) according to the manufacturer's instructions.

    Article Title: Brochothrix thermosphacta Bacteriophages Feature Heterogeneous and Highly Mosaic Genomes and Utilize Unique Prophage Insertion Sites ▿ Bacteriophages Feature Heterogeneous and Highly Mosaic Genomes and Utilize Unique Prophage Insertion Sites ▿ †
    Article Snippet: Phages were concentrated and purified by polyethylene glycol precipitation (PEG 8000, 7% in the presence of 1 M NaCl), stepped CsCl gradient ultracentrifugation , and dialysis against a 1,000-fold excess of SM buffer at 4°C. .. Bal31 nuclease treatment was performed by digestion of 40 μg of phage DNA with 20 U of Bal31 (New England Biolabs) at 30°C.

    Electrophoresis:

    Article Title: Brochothrix thermosphacta Bacteriophages Feature Heterogeneous and Highly Mosaic Genomes and Utilize Unique Prophage Insertion Sites ▿ Bacteriophages Feature Heterogeneous and Highly Mosaic Genomes and Utilize Unique Prophage Insertion Sites ▿ †
    Article Snippet: Bal31 nuclease treatment was performed by digestion of 40 μg of phage DNA with 20 U of Bal31 (New England Biolabs) at 30°C. .. An equal volume of each sample was digested with the indicated restriction enzyme, and products were electrophoresed in a GNA200 horizontal electrophoresis apparatus (GE Healthcare, Switzerland) at 2 V/cm in 1× Tris-acetate-EDTA buffer.

    Generated:

    Article Title: The mitochondrial genome of the pathogenic yeast Candida subhashii: GC-rich linear DNA with a protein covalently attached to the 5? termini
    Article Snippet: The terminal restriction enzyme fragments generated by Xba I endonuclease (1.5 and 2.8 kb) were cloned essentially as described for termini of linear mitochondrial plasmids of P. anserina ( ) and Pichia kluyveri ( ). .. Approximately 1 μg mtDNA was treated with either exonuclease III (New England Biolabs) or BAL-31 nuclease (New England Biolabs) according to the manufacturer's instructions.

    other:

    Article Title: Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles
    Article Snippet: To obtain preparations containing only covalently closed circles we eliminated unligated minicircles by the action of Bal31 nuclease.

    Article Title: Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles
    Article Snippet: Du et al . showed earlier that 63-bp DNA minicircles are so strongly bent that they form kinks and become sensitive to Bal31 nuclease, while 85-bp DNA minicircles can accommodate their bending stress without kinks and without becoming sensitive to Bal31 nuclease ( ).

    Article Title: Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles
    Article Snippet: However, torsionally relaxed extremely small DNA minicircles having only 63 base pairs were digested by Bal31 nuclease.

    Article Title: Production of DNA minicircles less than 250 base pairs through a novel concentrated DNA circularization assay enabling minicircle design with NF-κB inhibition activity
    Article Snippet: We next used Bal31 nuclease to detect the presence of DNA distortions within our constrained minicircles.

    Article Title: Cooperative kinking at distant sites in mechanically stressed DNA
    Article Snippet: This result indicates that the negative torsional stress sustained within MC100, previously shown to result in sensitivity to Bal31 nuclease, also leads to the creation of easily bendable sites that permit the molecules to minimize their bending energy by adopting elongated, elliptical shapes.

    Activity Assay:

    Article Title: The mitochondrial genome of the pathogenic yeast Candida subhashii: GC-rich linear DNA with a protein covalently attached to the 5? termini
    Article Snippet: Approximately 1 μg mtDNA was treated with either exonuclease III (New England Biolabs) or BAL-31 nuclease (New England Biolabs) according to the manufacturer's instructions. .. A linear 1040 bp long dsDNA fragment generated by digestion with Pvu II endonuclease was added to the C. subhashii mtDNA sample as an internal control of BAL-31 activity.

    Plasmid Preparation:

    Article Title: The mitochondrial genome of the pathogenic yeast Candida subhashii: GC-rich linear DNA with a protein covalently attached to the 5? termini
    Article Snippet: Next, Xba I-generated mtDNA fragments were ligated into the pUC19 vector digested with the endonucleases Sma I and Xba I. Sequences of five plasmid clones containing the terminal Xba I fragments were determined. .. Approximately 1 μg mtDNA was treated with either exonuclease III (New England Biolabs) or BAL-31 nuclease (New England Biolabs) according to the manufacturer's instructions.

    Sequencing:

    Article Title: Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles
    Article Snippet: .. However, the 94-bp-long minicircles with the sequence selected for their very efficient cyclization by C & W were not previously tested for their sensitivity to Bal31 nuclease. ..

    Molecular Weight:

    Article Title: Structural diversity of supercoiled DNA
    Article Snippet: .. BbvCI, EcoRV, Nb.BbvCI, NdeI, Nuclease Bal-31, T4 DNA Ligase, low molecular weight DNA ladder and 100 bp DNA ladder were purchased from New England Biolabs (Ipswich, MA). .. Proteinase K was purchased from Roche Molecular Biochemicals (Mannheim, Germany).

    DNA Sequencing:

    Article Title: The genome and proteome of Serratia bacteriophage ? which forms unstable lysogens
    Article Snippet: .. DNA sequencing and annotation Phage DNA was digested, ligated and treated with Bal31 exonuclease according to manufacturer’s specifications (NEB) and resolved using a 1% agarose gel. ..

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    New England Biolabs bal31 nuclease
    The formation and purification of 94-bp-long covalently closed DNA minicircles. A 2.5% agarose gel run in the presence of ethidium bromide (0.5 µg/ml) reveals that after the ligation reaction of minicircles with nicks, new species appear on the gel (compare lanes 3 with 2). Only these new species resist digestion by <t>Bal31</t> (lane 4) indicating that they are covalently closed DNA molecules. The electrophoretic migration of these new species is as expected for monomeric, dimeric and trimeric rings that acquired writhe due to ethidium bromide intercalation to covalently closed DNA molecules.
    Bal31 Nuclease, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bal31 nuclease/product/New England Biolabs
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    bal31 nuclease - by Bioz Stars, 2020-04
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    The formation and purification of 94-bp-long covalently closed DNA minicircles. A 2.5% agarose gel run in the presence of ethidium bromide (0.5 µg/ml) reveals that after the ligation reaction of minicircles with nicks, new species appear on the gel (compare lanes 3 with 2). Only these new species resist digestion by Bal31 (lane 4) indicating that they are covalently closed DNA molecules. The electrophoretic migration of these new species is as expected for monomeric, dimeric and trimeric rings that acquired writhe due to ethidium bromide intercalation to covalently closed DNA molecules.

    Journal: Nucleic Acids Research

    Article Title: Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles

    doi: 10.1093/nar/gkp137

    Figure Lengend Snippet: The formation and purification of 94-bp-long covalently closed DNA minicircles. A 2.5% agarose gel run in the presence of ethidium bromide (0.5 µg/ml) reveals that after the ligation reaction of minicircles with nicks, new species appear on the gel (compare lanes 3 with 2). Only these new species resist digestion by Bal31 (lane 4) indicating that they are covalently closed DNA molecules. The electrophoretic migration of these new species is as expected for monomeric, dimeric and trimeric rings that acquired writhe due to ethidium bromide intercalation to covalently closed DNA molecules.

    Article Snippet: However, the 94-bp-long minicircles with the sequence selected for their very efficient cyclization by C & W were not previously tested for their sensitivity to Bal31 nuclease.

    Techniques: Purification, Agarose Gel Electrophoresis, Ligation, Migration

    Bal31 nuclease detects a destabilized duplex DNA structure in covalently closed supercoiled DNA molecules but does not reveal the presence of kinks in 94-bp covalently closed DNA minicircles. A 2.5% agarose gel run in the presence of ethidium bromide (0.5 µg/ml) reveals that while negatively supercoiled DNA (lane 6) and nonligated nicked rings are completely digested by Bal31 nuclease (see lanes 2–4), the covalently closed monomeric, dimerc, trimeric and tetrameric circles (indicated with arrows) remain resistant to action of Bal31 nuclease (compare lanes 4 and 3).

    Journal: Nucleic Acids Research

    Article Title: Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles

    doi: 10.1093/nar/gkp137

    Figure Lengend Snippet: Bal31 nuclease detects a destabilized duplex DNA structure in covalently closed supercoiled DNA molecules but does not reveal the presence of kinks in 94-bp covalently closed DNA minicircles. A 2.5% agarose gel run in the presence of ethidium bromide (0.5 µg/ml) reveals that while negatively supercoiled DNA (lane 6) and nonligated nicked rings are completely digested by Bal31 nuclease (see lanes 2–4), the covalently closed monomeric, dimerc, trimeric and tetrameric circles (indicated with arrows) remain resistant to action of Bal31 nuclease (compare lanes 4 and 3).

    Article Snippet: However, the 94-bp-long minicircles with the sequence selected for their very efficient cyclization by C & W were not previously tested for their sensitivity to Bal31 nuclease.

    Techniques: Agarose Gel Electrophoresis

    DNA minicircles with 100, 106 and 108 bp, their purity and their susceptibility to Bal31 digestion. In a 10% denaturing PAGE gel (7 M urea), the denatured linear DNA fragments used as a marker, migrate much quicker than circular, covalently closed DNA minicircles in which the two strands cannot separate in space. Notice the presence of only one topoisomer in each category of minicircles and the sensitivity of minicircles with 100 bp to Bal31 nuclease.

    Journal: Nucleic Acids Research

    Article Title: Cooperative kinking at distant sites in mechanically stressed DNA

    doi: 10.1093/nar/gkr666

    Figure Lengend Snippet: DNA minicircles with 100, 106 and 108 bp, their purity and their susceptibility to Bal31 digestion. In a 10% denaturing PAGE gel (7 M urea), the denatured linear DNA fragments used as a marker, migrate much quicker than circular, covalently closed DNA minicircles in which the two strands cannot separate in space. Notice the presence of only one topoisomer in each category of minicircles and the sensitivity of minicircles with 100 bp to Bal31 nuclease.

    Article Snippet: This result indicates that the negative torsional stress sustained within MC100, previously shown to result in sensitivity to Bal31 nuclease, also leads to the creation of easily bendable sites that permit the molecules to minimize their bending energy by adopting elongated, elliptical shapes.

    Techniques: Polyacrylamide Gel Electrophoresis, Marker

    CNDCA-based production of supercoiled minicircle. Image of stained native polyacrylamide gel showing 95 bp minicircle topoisomers of decreased linking number (ΔLk values of −1 and −2) (lanes 4, 6) as indicated on the right and obtained by religation of the nicked minicircle in the presence of EtBr. The slowest migrating band corresponds to the relaxed minicircle that migrates expectedly at the same rate as the nicked minicircle (lanes 1 and 2). Digestion of minicircle samples by the nuclease Bal31 is indicated on the right. The position of the molecular mass markers in bp is indicated on the left.

    Journal: Nucleic Acids Research

    Article Title: Production of DNA minicircles less than 250 base pairs through a novel concentrated DNA circularization assay enabling minicircle design with NF-κB inhibition activity

    doi: 10.1093/nar/gkw1034

    Figure Lengend Snippet: CNDCA-based production of supercoiled minicircle. Image of stained native polyacrylamide gel showing 95 bp minicircle topoisomers of decreased linking number (ΔLk values of −1 and −2) (lanes 4, 6) as indicated on the right and obtained by religation of the nicked minicircle in the presence of EtBr. The slowest migrating band corresponds to the relaxed minicircle that migrates expectedly at the same rate as the nicked minicircle (lanes 1 and 2). Digestion of minicircle samples by the nuclease Bal31 is indicated on the right. The position of the molecular mass markers in bp is indicated on the left.

    Article Snippet: We next used Bal31 nuclease to detect the presence of DNA distortions within our constrained minicircles.

    Techniques: Staining