bsa ji  (New England Biolabs)


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    New England Biolabs bsa ji
    p53 codon 72 genotyping . (A) PCR for TP53 exon 4 detection. PCR products were loaded on 1.5% agarose gel. 100 base pairs DNA ladder (lane 1), positive controls (lane 2 and 3), negative controls (lanes 4 and 26), samples tested (lanes 5-25). (B) Detection of TP53 codon 72 polymorphism by DHPLC. Elution profiles obtained at 59°C. Representative heterozygous (a) and homozygous profile (b and c) are depicted. Note the small shoulder only in heterozygous samples. After the first trial, homozygous samples were mixed in approximately equimolar proportions with a control sample with TP53 codon 72 polymorphism previously identified as homozygous proline. This allowed differentiating the two homozygous genotypes. Homozygous profiles represent a sample with the genotype similar to the control sample added. Heterozygous profile represents a sample with the genotype differing from the control sample. (C) Dot Blot hybridization for TP53 codon 72 polymorphism genotyping. Samples were spotted onto nylon membranes and hybridized with biotin-labelled oligonucleotide probes for allele Arg (PArg) and allele Pro (PPro). Sample 1A, 1B and 2A represent positive controls. The profile observed in sample 1A is compatible with a homozygous arginine. Sample 1B presents a profile compatible with the genotype homozygous proline. Sample 2A presents a profile compatible with the heterozygous genotype. (D) Silver-stained 8% polyacrylamide gel showing the restriction profiles obtained with enzyme <t>Bst</t> UI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 2, 4, 5 represent heterozygous samples where fragments of 279, 160 and 119 base pairs can be detected. Lanes 3 and 8 represent homozygous proline samples in which the enzyme was not able to digest the PCR product. Lanes 6 represent an homozygous arginine in which fragments of 160 and 119 base pairs can be identified. Lanes 7, 9 and 10 represent positive controls for homozygous arginine, homozygous proline and heterozygous samples, respectively. (E) Silver-stained 12% polyacrylamide gel showing the restriction profiles obtained with enzyme <t>Bsa</t> JI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 3, 4, 5 represent heterozygous samples in which fragments of 138, 94, 66, 44, 41 and 34 base pairs can be detected. Lanes 7 represent a homozygous proline sample in which fragments of 94, 66, 44, 41 and 34 base pairs can be identified. Lanes 2, 6, 8, 10 and 11 represent homozygous arginine samples in which fragments of 138, 66, 41 and 34 base pairs can be detected. Lanes 9, 12 and 13 represent positive controls for heterozygous, homozygous arginine and homozygous proline samples, respectively.
    Bsa Ji, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsa ji/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bsa ji - by Bioz Stars, 2021-04
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    1) Product Images from "Validation of dot blot hybridization and denaturing high performance liquid chromatography as reliable methods for TP53 codon 72 genotyping in molecular epidemiologic studies"

    Article Title: Validation of dot blot hybridization and denaturing high performance liquid chromatography as reliable methods for TP53 codon 72 genotyping in molecular epidemiologic studies

    Journal: BMC Genetics

    doi: 10.1186/1471-2156-11-44

    p53 codon 72 genotyping . (A) PCR for TP53 exon 4 detection. PCR products were loaded on 1.5% agarose gel. 100 base pairs DNA ladder (lane 1), positive controls (lane 2 and 3), negative controls (lanes 4 and 26), samples tested (lanes 5-25). (B) Detection of TP53 codon 72 polymorphism by DHPLC. Elution profiles obtained at 59°C. Representative heterozygous (a) and homozygous profile (b and c) are depicted. Note the small shoulder only in heterozygous samples. After the first trial, homozygous samples were mixed in approximately equimolar proportions with a control sample with TP53 codon 72 polymorphism previously identified as homozygous proline. This allowed differentiating the two homozygous genotypes. Homozygous profiles represent a sample with the genotype similar to the control sample added. Heterozygous profile represents a sample with the genotype differing from the control sample. (C) Dot Blot hybridization for TP53 codon 72 polymorphism genotyping. Samples were spotted onto nylon membranes and hybridized with biotin-labelled oligonucleotide probes for allele Arg (PArg) and allele Pro (PPro). Sample 1A, 1B and 2A represent positive controls. The profile observed in sample 1A is compatible with a homozygous arginine. Sample 1B presents a profile compatible with the genotype homozygous proline. Sample 2A presents a profile compatible with the heterozygous genotype. (D) Silver-stained 8% polyacrylamide gel showing the restriction profiles obtained with enzyme Bst UI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 2, 4, 5 represent heterozygous samples where fragments of 279, 160 and 119 base pairs can be detected. Lanes 3 and 8 represent homozygous proline samples in which the enzyme was not able to digest the PCR product. Lanes 6 represent an homozygous arginine in which fragments of 160 and 119 base pairs can be identified. Lanes 7, 9 and 10 represent positive controls for homozygous arginine, homozygous proline and heterozygous samples, respectively. (E) Silver-stained 12% polyacrylamide gel showing the restriction profiles obtained with enzyme Bsa JI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 3, 4, 5 represent heterozygous samples in which fragments of 138, 94, 66, 44, 41 and 34 base pairs can be detected. Lanes 7 represent a homozygous proline sample in which fragments of 94, 66, 44, 41 and 34 base pairs can be identified. Lanes 2, 6, 8, 10 and 11 represent homozygous arginine samples in which fragments of 138, 66, 41 and 34 base pairs can be detected. Lanes 9, 12 and 13 represent positive controls for heterozygous, homozygous arginine and homozygous proline samples, respectively.
    Figure Legend Snippet: p53 codon 72 genotyping . (A) PCR for TP53 exon 4 detection. PCR products were loaded on 1.5% agarose gel. 100 base pairs DNA ladder (lane 1), positive controls (lane 2 and 3), negative controls (lanes 4 and 26), samples tested (lanes 5-25). (B) Detection of TP53 codon 72 polymorphism by DHPLC. Elution profiles obtained at 59°C. Representative heterozygous (a) and homozygous profile (b and c) are depicted. Note the small shoulder only in heterozygous samples. After the first trial, homozygous samples were mixed in approximately equimolar proportions with a control sample with TP53 codon 72 polymorphism previously identified as homozygous proline. This allowed differentiating the two homozygous genotypes. Homozygous profiles represent a sample with the genotype similar to the control sample added. Heterozygous profile represents a sample with the genotype differing from the control sample. (C) Dot Blot hybridization for TP53 codon 72 polymorphism genotyping. Samples were spotted onto nylon membranes and hybridized with biotin-labelled oligonucleotide probes for allele Arg (PArg) and allele Pro (PPro). Sample 1A, 1B and 2A represent positive controls. The profile observed in sample 1A is compatible with a homozygous arginine. Sample 1B presents a profile compatible with the genotype homozygous proline. Sample 2A presents a profile compatible with the heterozygous genotype. (D) Silver-stained 8% polyacrylamide gel showing the restriction profiles obtained with enzyme Bst UI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 2, 4, 5 represent heterozygous samples where fragments of 279, 160 and 119 base pairs can be detected. Lanes 3 and 8 represent homozygous proline samples in which the enzyme was not able to digest the PCR product. Lanes 6 represent an homozygous arginine in which fragments of 160 and 119 base pairs can be identified. Lanes 7, 9 and 10 represent positive controls for homozygous arginine, homozygous proline and heterozygous samples, respectively. (E) Silver-stained 12% polyacrylamide gel showing the restriction profiles obtained with enzyme Bsa JI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 3, 4, 5 represent heterozygous samples in which fragments of 138, 94, 66, 44, 41 and 34 base pairs can be detected. Lanes 7 represent a homozygous proline sample in which fragments of 94, 66, 44, 41 and 34 base pairs can be identified. Lanes 2, 6, 8, 10 and 11 represent homozygous arginine samples in which fragments of 138, 66, 41 and 34 base pairs can be detected. Lanes 9, 12 and 13 represent positive controls for heterozygous, homozygous arginine and homozygous proline samples, respectively.

    Techniques Used: Polymerase Chain Reaction, Agarose Gel Electrophoresis, Dot Blot, Hybridization, Staining

    Related Articles

    GWAS:

    Article Title: Cyclic AMP and c-KIT Signaling in Familial Testicular Germ Cell Tumor Predisposition
    Article Snippet: Because both the University of Pennsylvania study ( ) and our own familial cohort consisted entirely of US white men, the GWAS KITLG single-nucleotide polymorphisms (SNPs) for the current analysis were selected from that GWAS. .. The presence of the KITLG variants that were most strongly associated with TGCT risk in the recently published GWAS , rs3782179 and rs4474514, was investigated in patients with FTGCT and their family members using restriction enzymes Bsa JI and Bgl II (New England Biolabs Inc, Ipswich, Massachusetts), respectively; when positive, their presence was confirmed by sequencing. .. The 692 New York Cancer Project samples were then genotyped using the Mass-Array matrix-assisted laser desorption ionization time-of-flight mass spectrometry-based SNP genotyping system, according to the manufacturer's protocol (Sequenom, San Diego, California), as previously published ( ).

    Sequencing:

    Article Title: Cyclic AMP and c-KIT Signaling in Familial Testicular Germ Cell Tumor Predisposition
    Article Snippet: Because both the University of Pennsylvania study ( ) and our own familial cohort consisted entirely of US white men, the GWAS KITLG single-nucleotide polymorphisms (SNPs) for the current analysis were selected from that GWAS. .. The presence of the KITLG variants that were most strongly associated with TGCT risk in the recently published GWAS , rs3782179 and rs4474514, was investigated in patients with FTGCT and their family members using restriction enzymes Bsa JI and Bgl II (New England Biolabs Inc, Ipswich, Massachusetts), respectively; when positive, their presence was confirmed by sequencing. .. The 692 New York Cancer Project samples were then genotyped using the Mass-Array matrix-assisted laser desorption ionization time-of-flight mass spectrometry-based SNP genotyping system, according to the manufacturer's protocol (Sequenom, San Diego, California), as previously published ( ).

    Incubation:

    Article Title: The nitrosated bile acid DNA lesion O6-carboxymethylguanine is a substrate for the human DNA repair protein O6-methylguanine-DNA methyltransferase
    Article Snippet: .. The resulting double-stranded ODNs (7 pmoles) were incubated with 21 pmoles MGMT in a total volume of 17 µl of IBSA (Buffer I containing 0.1% BSA) at room temperature for 3 h and then subjected to digestion with NlaIII, CviKI, StyI, BsaJI or PstI in the buffer provided by the manufacturer (New England Biolabs) in a total volume of 10 µl at room temperature for 1 h. Non-denaturing PAGE gel-loading buffer (30% glycerol, 0.25% xylene cyanol, 0.25% bromophenol blue: 2 µl) was added to each sample, which was then applied to a 20% PAGE gel and subjected to electrophoresis at 100 volts for 1 h. The gels were scanned on a Pharos phosphorimager (BioRad) using the fluorescent gel scanner settings. .. Inhibition of MGMT by O6 -alkG-containing ODNs Duplex ODNs containing O 6 -MeG and O 6 -CMG, but not the control ODN (which contained G in place of the alkylated base), potently inhibited the action of MGMT on [3 H]-methylated substrate DNA.

    Polyacrylamide Gel Electrophoresis:

    Article Title: The nitrosated bile acid DNA lesion O6-carboxymethylguanine is a substrate for the human DNA repair protein O6-methylguanine-DNA methyltransferase
    Article Snippet: .. The resulting double-stranded ODNs (7 pmoles) were incubated with 21 pmoles MGMT in a total volume of 17 µl of IBSA (Buffer I containing 0.1% BSA) at room temperature for 3 h and then subjected to digestion with NlaIII, CviKI, StyI, BsaJI or PstI in the buffer provided by the manufacturer (New England Biolabs) in a total volume of 10 µl at room temperature for 1 h. Non-denaturing PAGE gel-loading buffer (30% glycerol, 0.25% xylene cyanol, 0.25% bromophenol blue: 2 µl) was added to each sample, which was then applied to a 20% PAGE gel and subjected to electrophoresis at 100 volts for 1 h. The gels were scanned on a Pharos phosphorimager (BioRad) using the fluorescent gel scanner settings. .. Inhibition of MGMT by O6 -alkG-containing ODNs Duplex ODNs containing O 6 -MeG and O 6 -CMG, but not the control ODN (which contained G in place of the alkylated base), potently inhibited the action of MGMT on [3 H]-methylated substrate DNA.

    Electrophoresis:

    Article Title: The nitrosated bile acid DNA lesion O6-carboxymethylguanine is a substrate for the human DNA repair protein O6-methylguanine-DNA methyltransferase
    Article Snippet: .. The resulting double-stranded ODNs (7 pmoles) were incubated with 21 pmoles MGMT in a total volume of 17 µl of IBSA (Buffer I containing 0.1% BSA) at room temperature for 3 h and then subjected to digestion with NlaIII, CviKI, StyI, BsaJI or PstI in the buffer provided by the manufacturer (New England Biolabs) in a total volume of 10 µl at room temperature for 1 h. Non-denaturing PAGE gel-loading buffer (30% glycerol, 0.25% xylene cyanol, 0.25% bromophenol blue: 2 µl) was added to each sample, which was then applied to a 20% PAGE gel and subjected to electrophoresis at 100 volts for 1 h. The gels were scanned on a Pharos phosphorimager (BioRad) using the fluorescent gel scanner settings. .. Inhibition of MGMT by O6 -alkG-containing ODNs Duplex ODNs containing O 6 -MeG and O 6 -CMG, but not the control ODN (which contained G in place of the alkylated base), potently inhibited the action of MGMT on [3 H]-methylated substrate DNA.

    Polymerase Chain Reaction:

    Article Title: Mutation of melanosome protein RAB38 in chocolate mice
    Article Snippet: Mutations were confirmed by using primers designed to amplify a 213-bp fragment surrounding the G146T sequence ( cht Ex1F-GGCCTCCAGGATGCAGACACC and cht Ex1R-CCAGCAATGTCCCAGAGCTGC). .. Sex AI and Bsa JI restriction enzyme digests were performed by using 10 μl of PCR product, 2.5 units enzyme (New England Biolabs) along with 1× of the supplied BSA and digest buffer. ..

    Article Title: Efficient CRISPR/Cas9-mediated biallelic gene disruption and site-specific knockin after rapid selection of highly active sgRNAs in pigs
    Article Snippet: The identity of each PCR product was confirmed by restriction fragment length polymorphism (RFLP) analysis. .. Ten microliters of PCR product was digested with BsaJI, HpyCH4V or DraI (New England Biolabs, US), respectively. .. The reaction products were separated by 2% agarose gel electrophoresis in the presence of ethidium bromide solution, and visualized with a UV transilluminator (UVP, Upland, CA).

    Amplification:

    Article Title: Validation of dot blot hybridization and denaturing high performance liquid chromatography as reliable methods for TP53 codon 72 genotyping in molecular epidemiologic studies
    Article Snippet: If needed to obtain optimal signal intensities, the exposure times were sometimes varied, as judged by evaluation of the hybridization controls present on each membrane. .. Restriction Fragment Length Polymorphism (RFLP) After TP53 exon 4 amplification, 7 μL of the amplified reaction products were digested in a 10 μL volume of 1 × NEB buffer 2 and 5 U of Bsa JI or Bst UI endonucleases (New England BioLabs, Newton, MA) at 60°C for at least 2 hours. ..

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    • bsa ji  (New England Biolabs)
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    New England Biolabs bsa ji
    p53 codon 72 genotyping . (A) PCR for TP53 exon 4 detection. PCR products were loaded on 1.5% agarose gel. 100 base pairs DNA ladder (lane 1), positive controls (lane 2 and 3), negative controls (lanes 4 and 26), samples tested (lanes 5-25). (B) Detection of TP53 codon 72 polymorphism by DHPLC. Elution profiles obtained at 59°C. Representative heterozygous (a) and homozygous profile (b and c) are depicted. Note the small shoulder only in heterozygous samples. After the first trial, homozygous samples were mixed in approximately equimolar proportions with a control sample with TP53 codon 72 polymorphism previously identified as homozygous proline. This allowed differentiating the two homozygous genotypes. Homozygous profiles represent a sample with the genotype similar to the control sample added. Heterozygous profile represents a sample with the genotype differing from the control sample. (C) Dot Blot hybridization for TP53 codon 72 polymorphism genotyping. Samples were spotted onto nylon membranes and hybridized with biotin-labelled oligonucleotide probes for allele Arg (PArg) and allele Pro (PPro). Sample 1A, 1B and 2A represent positive controls. The profile observed in sample 1A is compatible with a homozygous arginine. Sample 1B presents a profile compatible with the genotype homozygous proline. Sample 2A presents a profile compatible with the heterozygous genotype. (D) Silver-stained 8% polyacrylamide gel showing the restriction profiles obtained with enzyme <t>Bst</t> UI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 2, 4, 5 represent heterozygous samples where fragments of 279, 160 and 119 base pairs can be detected. Lanes 3 and 8 represent homozygous proline samples in which the enzyme was not able to digest the PCR product. Lanes 6 represent an homozygous arginine in which fragments of 160 and 119 base pairs can be identified. Lanes 7, 9 and 10 represent positive controls for homozygous arginine, homozygous proline and heterozygous samples, respectively. (E) Silver-stained 12% polyacrylamide gel showing the restriction profiles obtained with enzyme <t>Bsa</t> JI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 3, 4, 5 represent heterozygous samples in which fragments of 138, 94, 66, 44, 41 and 34 base pairs can be detected. Lanes 7 represent a homozygous proline sample in which fragments of 94, 66, 44, 41 and 34 base pairs can be identified. Lanes 2, 6, 8, 10 and 11 represent homozygous arginine samples in which fragments of 138, 66, 41 and 34 base pairs can be detected. Lanes 9, 12 and 13 represent positive controls for heterozygous, homozygous arginine and homozygous proline samples, respectively.
    Bsa Ji, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bsa ji/product/New England Biolabs
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    bsa ji - by Bioz Stars, 2021-04
    86/100 stars
    		  Buy from Supplier

    Image Search Results


    p53 codon 72 genotyping . (A) PCR for TP53 exon 4 detection. PCR products were loaded on 1.5% agarose gel. 100 base pairs DNA ladder (lane 1), positive controls (lane 2 and 3), negative controls (lanes 4 and 26), samples tested (lanes 5-25). (B) Detection of TP53 codon 72 polymorphism by DHPLC. Elution profiles obtained at 59°C. Representative heterozygous (a) and homozygous profile (b and c) are depicted. Note the small shoulder only in heterozygous samples. After the first trial, homozygous samples were mixed in approximately equimolar proportions with a control sample with TP53 codon 72 polymorphism previously identified as homozygous proline. This allowed differentiating the two homozygous genotypes. Homozygous profiles represent a sample with the genotype similar to the control sample added. Heterozygous profile represents a sample with the genotype differing from the control sample. (C) Dot Blot hybridization for TP53 codon 72 polymorphism genotyping. Samples were spotted onto nylon membranes and hybridized with biotin-labelled oligonucleotide probes for allele Arg (PArg) and allele Pro (PPro). Sample 1A, 1B and 2A represent positive controls. The profile observed in sample 1A is compatible with a homozygous arginine. Sample 1B presents a profile compatible with the genotype homozygous proline. Sample 2A presents a profile compatible with the heterozygous genotype. (D) Silver-stained 8% polyacrylamide gel showing the restriction profiles obtained with enzyme Bst UI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 2, 4, 5 represent heterozygous samples where fragments of 279, 160 and 119 base pairs can be detected. Lanes 3 and 8 represent homozygous proline samples in which the enzyme was not able to digest the PCR product. Lanes 6 represent an homozygous arginine in which fragments of 160 and 119 base pairs can be identified. Lanes 7, 9 and 10 represent positive controls for homozygous arginine, homozygous proline and heterozygous samples, respectively. (E) Silver-stained 12% polyacrylamide gel showing the restriction profiles obtained with enzyme Bsa JI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 3, 4, 5 represent heterozygous samples in which fragments of 138, 94, 66, 44, 41 and 34 base pairs can be detected. Lanes 7 represent a homozygous proline sample in which fragments of 94, 66, 44, 41 and 34 base pairs can be identified. Lanes 2, 6, 8, 10 and 11 represent homozygous arginine samples in which fragments of 138, 66, 41 and 34 base pairs can be detected. Lanes 9, 12 and 13 represent positive controls for heterozygous, homozygous arginine and homozygous proline samples, respectively.

    Journal: BMC Genetics

    Article Title: Validation of dot blot hybridization and denaturing high performance liquid chromatography as reliable methods for TP53 codon 72 genotyping in molecular epidemiologic studies

    doi: 10.1186/1471-2156-11-44

    Figure Lengend Snippet: p53 codon 72 genotyping . (A) PCR for TP53 exon 4 detection. PCR products were loaded on 1.5% agarose gel. 100 base pairs DNA ladder (lane 1), positive controls (lane 2 and 3), negative controls (lanes 4 and 26), samples tested (lanes 5-25). (B) Detection of TP53 codon 72 polymorphism by DHPLC. Elution profiles obtained at 59°C. Representative heterozygous (a) and homozygous profile (b and c) are depicted. Note the small shoulder only in heterozygous samples. After the first trial, homozygous samples were mixed in approximately equimolar proportions with a control sample with TP53 codon 72 polymorphism previously identified as homozygous proline. This allowed differentiating the two homozygous genotypes. Homozygous profiles represent a sample with the genotype similar to the control sample added. Heterozygous profile represents a sample with the genotype differing from the control sample. (C) Dot Blot hybridization for TP53 codon 72 polymorphism genotyping. Samples were spotted onto nylon membranes and hybridized with biotin-labelled oligonucleotide probes for allele Arg (PArg) and allele Pro (PPro). Sample 1A, 1B and 2A represent positive controls. The profile observed in sample 1A is compatible with a homozygous arginine. Sample 1B presents a profile compatible with the genotype homozygous proline. Sample 2A presents a profile compatible with the heterozygous genotype. (D) Silver-stained 8% polyacrylamide gel showing the restriction profiles obtained with enzyme Bst UI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 2, 4, 5 represent heterozygous samples where fragments of 279, 160 and 119 base pairs can be detected. Lanes 3 and 8 represent homozygous proline samples in which the enzyme was not able to digest the PCR product. Lanes 6 represent an homozygous arginine in which fragments of 160 and 119 base pairs can be identified. Lanes 7, 9 and 10 represent positive controls for homozygous arginine, homozygous proline and heterozygous samples, respectively. (E) Silver-stained 12% polyacrylamide gel showing the restriction profiles obtained with enzyme Bsa JI of the TP53 exon 4 PCR product. 50 base pairs DNA ladder (lane 1). Lanes 3, 4, 5 represent heterozygous samples in which fragments of 138, 94, 66, 44, 41 and 34 base pairs can be detected. Lanes 7 represent a homozygous proline sample in which fragments of 94, 66, 44, 41 and 34 base pairs can be identified. Lanes 2, 6, 8, 10 and 11 represent homozygous arginine samples in which fragments of 138, 66, 41 and 34 base pairs can be detected. Lanes 9, 12 and 13 represent positive controls for heterozygous, homozygous arginine and homozygous proline samples, respectively.

    Article Snippet: Restriction Fragment Length Polymorphism (RFLP) After TP53 exon 4 amplification, 7 μL of the amplified reaction products were digested in a 10 μL volume of 1 × NEB buffer 2 and 5 U of Bsa JI or Bst UI endonucleases (New England BioLabs, Newton, MA) at 60°C for at least 2 hours.

    Techniques: Polymerase Chain Reaction, Agarose Gel Electrophoresis, Dot Blot, Hybridization, Staining