hpych4v  (New England Biolabs)


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    Name:
    HpyCH4V
    Description:
    HpyCH4V 500 units
    Catalog Number:
    R0620L
    Price:
    282
    Category:
    Restriction Enzymes
    Size:
    500 units
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    Structured Review

    New England Biolabs hpych4v
    HpyCH4V
    HpyCH4V 500 units
    https://www.bioz.com/result/hpych4v/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hpych4v - by Bioz Stars, 2021-06
    95/100 stars

    Images

    1) Product Images from "Seven novel mutations in the long isoform of the USH2A gene in Chinese families with nonsyndromic retinitis pigmentosa and Usher syndrome Type II"

    Article Title: Seven novel mutations in the long isoform of the USH2A gene in Chinese families with nonsyndromic retinitis pigmentosa and Usher syndrome Type II

    Journal: Molecular Vision

    doi:

    A restriction fragment length analysis of the four mutations detected in this study. A : c.2802T > G abolished a HincII restriction site that co-segregated with the affected individuals and the carriers (42 bp, 57 bp, 99 bp, 717 bp, and 774 bp), but not with unaffected individuals and normal controls (42 bp, 57 bp, and 717 bp). B : c.8232G > C created a new HpyCH4V restriction site that co-segregated with the affected individuals and the carriers (88 bp, 186 bp, 218 bp, and 274 bp), but not with unaffected individuals and normal controls (218 bp, 274 bp). C : c.3788G > A abolished a BsaI restriction site that co-segregated with the affected individuals and the carriers (70 bp, 132 bp, 422 bp, and 492 bp), but not with unaffected individuals and normal controls (70 bp, 132 bp, and 422 bp). D : c.14403C > G created a SpeI restriction site that co-segregated with the affected individuals and the carriers (145 bp, 300 bp, and 445 bp), but not with unaffected individuals and normal controls (445 bp). A participant identification number is given above each lane. N represents normal controls.
    Figure Legend Snippet: A restriction fragment length analysis of the four mutations detected in this study. A : c.2802T > G abolished a HincII restriction site that co-segregated with the affected individuals and the carriers (42 bp, 57 bp, 99 bp, 717 bp, and 774 bp), but not with unaffected individuals and normal controls (42 bp, 57 bp, and 717 bp). B : c.8232G > C created a new HpyCH4V restriction site that co-segregated with the affected individuals and the carriers (88 bp, 186 bp, 218 bp, and 274 bp), but not with unaffected individuals and normal controls (218 bp, 274 bp). C : c.3788G > A abolished a BsaI restriction site that co-segregated with the affected individuals and the carriers (70 bp, 132 bp, 422 bp, and 492 bp), but not with unaffected individuals and normal controls (70 bp, 132 bp, and 422 bp). D : c.14403C > G created a SpeI restriction site that co-segregated with the affected individuals and the carriers (145 bp, 300 bp, and 445 bp), but not with unaffected individuals and normal controls (445 bp). A participant identification number is given above each lane. N represents normal controls.

    Techniques Used:

    2) Product Images from "Efficient CRISPR/Cas9-mediated biallelic gene disruption and site-specific knockin after rapid selection of highly active sgRNAs in pigs"

    Article Title: Efficient CRISPR/Cas9-mediated biallelic gene disruption and site-specific knockin after rapid selection of highly active sgRNAs in pigs

    Journal: Scientific Reports

    doi: 10.1038/srep13348

    Rapidly selecting most effective sgRNAs by single blastocyst genotyping. ( A ) Overview and timeline of the experiment. Beginning wit h experimental design, assessment of sgRNA mutagenesis efficiencies can be achieved within 10 days. ( B ) Genotyping of single blastocyst derived from Cas9 mRNA and sgRNA-injected parthenogenetic oocytes by RFLP and Sanger sequence analyses. Representative RFLP agarose gel electrophoresis showing PCR product of target region derived from 10 individual blastocysts digested with different restriction enzymes. The F1, F2, R1 and R2 sgRNA mutagenesis efficiencies were assessed by BsaJI, HpyCH4V, BsaJI and DraI, respectively. In each RFLP assay, some PCR products were sequenced to confirm the mutations at the target sites. The numbers on the right show the type of mutation and how many nucleotides are involved, with “−” and “+” indicating deletion or insertion of the given number of nucleotides, respectively. The sgRNA sequence is labeled in red, and the PAM sequence is labeled in purple. Deleted bases are marked with colons, and arrows indicate the sites of inserted bases, which are listed under the mutant alleles.
    Figure Legend Snippet: Rapidly selecting most effective sgRNAs by single blastocyst genotyping. ( A ) Overview and timeline of the experiment. Beginning wit h experimental design, assessment of sgRNA mutagenesis efficiencies can be achieved within 10 days. ( B ) Genotyping of single blastocyst derived from Cas9 mRNA and sgRNA-injected parthenogenetic oocytes by RFLP and Sanger sequence analyses. Representative RFLP agarose gel electrophoresis showing PCR product of target region derived from 10 individual blastocysts digested with different restriction enzymes. The F1, F2, R1 and R2 sgRNA mutagenesis efficiencies were assessed by BsaJI, HpyCH4V, BsaJI and DraI, respectively. In each RFLP assay, some PCR products were sequenced to confirm the mutations at the target sites. The numbers on the right show the type of mutation and how many nucleotides are involved, with “−” and “+” indicating deletion or insertion of the given number of nucleotides, respectively. The sgRNA sequence is labeled in red, and the PAM sequence is labeled in purple. Deleted bases are marked with colons, and arrows indicate the sites of inserted bases, which are listed under the mutant alleles.

    Techniques Used: Mutagenesis, Derivative Assay, Injection, Sequencing, Agarose Gel Electrophoresis, Polymerase Chain Reaction, RFLP Assay, Labeling

    3) Product Images from "Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification"

    Article Title: Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification

    Journal: Journal of Virological Methods

    doi: 10.1016/j.jviromet.2015.02.001

    Restriction enzyme digestion of RT-LAMP products. Undigested RT-LAMP products (Lane 1) and products digested with HpyCH4V (Lane 2). M, marker.
    Figure Legend Snippet: Restriction enzyme digestion of RT-LAMP products. Undigested RT-LAMP products (Lane 1) and products digested with HpyCH4V (Lane 2). M, marker.

    Techniques Used: Marker

    4) Product Images from "Novel pathogenic mutations in C1QTNF5 support a dominant negative disease mechanism in late-onset retinal degeneration"

    Article Title: Novel pathogenic mutations in C1QTNF5 support a dominant negative disease mechanism in late-onset retinal degeneration

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-11898-3

    Pedigree, genotype and phenotype of family 1. ( a ) L-ORD family 1 is a multi-generation pedigree showing autosomal dominant inheritance. Squares represent males, circles represent females. Shaded shapes indicate an affected individual. Clinical phenotyping of the proband V:I. ( b ) Anterior segment slit lamp photograph shows long-anterior zonules (white arrow) encroaching beyond the pupil margin in a dilated pupil. ( c ) Colour fundus photography shows areas of central atrophy affecting the macula and ( d ) Fundus autofluorescence imaging shows areas of central scalloped hypoautofluorescence matching the areas of atrophy surrounded by a cuff of increased autofluorescence. ( e ) Microperimetry interpolated plot superimposes areas of retinal sensitivity on a colour fundus image. Areas of good sensitvity vary from green to yellow to red for poor sensitivity. Areas with no sensitivity have no polygons. The image shows non-detectable sensitivity at the macula for the patient. ( f ) Hyperreflective deposits deep to the neural retina, indicated by asterisks, are observed in the macula in an OCT scan of the proband. NR, neural retina; CH, choroid. ( g ) Dark adaptometry of the proband (L-ORD case) shows that recovery of sensitivity after light exposure is slow and final sensitivity is still reduced at 80 minutes, compared to the normal, rapid recovery of sensitivity in an unaffected sibling. ( h ) Sequence analysis of C1QTNF5 exon 3 revealed a heterozygous c.646G > T transversion, resulting in a glycine to cysteine mutation at position 216 in the L-ORD case (lower panel) compared to an unaffected relative (upper panel). ( i ) HpyCh4V restriction digest confirms the c.646 G > T mutation by generating fragments of 439, 363, 129 and 120 bp in homozygous G (p.G216) samples, and 439, 363, 129, 94 and 26 bp for the mutant G > T (p.C216) allele.
    Figure Legend Snippet: Pedigree, genotype and phenotype of family 1. ( a ) L-ORD family 1 is a multi-generation pedigree showing autosomal dominant inheritance. Squares represent males, circles represent females. Shaded shapes indicate an affected individual. Clinical phenotyping of the proband V:I. ( b ) Anterior segment slit lamp photograph shows long-anterior zonules (white arrow) encroaching beyond the pupil margin in a dilated pupil. ( c ) Colour fundus photography shows areas of central atrophy affecting the macula and ( d ) Fundus autofluorescence imaging shows areas of central scalloped hypoautofluorescence matching the areas of atrophy surrounded by a cuff of increased autofluorescence. ( e ) Microperimetry interpolated plot superimposes areas of retinal sensitivity on a colour fundus image. Areas of good sensitvity vary from green to yellow to red for poor sensitivity. Areas with no sensitivity have no polygons. The image shows non-detectable sensitivity at the macula for the patient. ( f ) Hyperreflective deposits deep to the neural retina, indicated by asterisks, are observed in the macula in an OCT scan of the proband. NR, neural retina; CH, choroid. ( g ) Dark adaptometry of the proband (L-ORD case) shows that recovery of sensitivity after light exposure is slow and final sensitivity is still reduced at 80 minutes, compared to the normal, rapid recovery of sensitivity in an unaffected sibling. ( h ) Sequence analysis of C1QTNF5 exon 3 revealed a heterozygous c.646G > T transversion, resulting in a glycine to cysteine mutation at position 216 in the L-ORD case (lower panel) compared to an unaffected relative (upper panel). ( i ) HpyCh4V restriction digest confirms the c.646 G > T mutation by generating fragments of 439, 363, 129 and 120 bp in homozygous G (p.G216) samples, and 439, 363, 129, 94 and 26 bp for the mutant G > T (p.C216) allele.

    Techniques Used: Imaging, Sequencing, Mutagenesis

    5) Product Images from "Novel pathogenic mutations in C1QTNF5 support a dominant negative disease mechanism in late-onset retinal degeneration"

    Article Title: Novel pathogenic mutations in C1QTNF5 support a dominant negative disease mechanism in late-onset retinal degeneration

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-11898-3

    Pedigree, genotype and phenotype of family 1. ( a ) L-ORD family 1 is a multi-generation pedigree showing autosomal dominant inheritance. Squares represent males, circles represent females. Shaded shapes indicate an affected individual. Clinical phenotyping of the proband V:I. ( b ) Anterior segment slit lamp photograph shows long-anterior zonules (white arrow) encroaching beyond the pupil margin in a dilated pupil. ( c ) Colour fundus photography shows areas of central atrophy affecting the macula and ( d ) Fundus autofluorescence imaging shows areas of central scalloped hypoautofluorescence matching the areas of atrophy surrounded by a cuff of increased autofluorescence. ( e ) Microperimetry interpolated plot superimposes areas of retinal sensitivity on a colour fundus image. Areas of good sensitvity vary from green to yellow to red for poor sensitivity. Areas with no sensitivity have no polygons. The image shows non-detectable sensitivity at the macula for the patient. ( f ) Hyperreflective deposits deep to the neural retina, indicated by asterisks, are observed in the macula in an OCT scan of the proband. NR, neural retina; CH, choroid. ( g ) Dark adaptometry of the proband (L-ORD case) shows that recovery of sensitivity after light exposure is slow and final sensitivity is still reduced at 80 minutes, compared to the normal, rapid recovery of sensitivity in an unaffected sibling. ( h ) Sequence analysis of C1QTNF5 exon 3 revealed a heterozygous c.646G > T transversion, resulting in a glycine to cysteine mutation at position 216 in the L-ORD case (lower panel) compared to an unaffected relative (upper panel). ( i ) HpyCh4V restriction digest confirms the c.646 G > T mutation by generating fragments of 439, 363, 129 and 120 bp in homozygous G (p.G216) samples, and 439, 363, 129, 94 and 26 bp for the mutant G > T (p.C216) allele.
    Figure Legend Snippet: Pedigree, genotype and phenotype of family 1. ( a ) L-ORD family 1 is a multi-generation pedigree showing autosomal dominant inheritance. Squares represent males, circles represent females. Shaded shapes indicate an affected individual. Clinical phenotyping of the proband V:I. ( b ) Anterior segment slit lamp photograph shows long-anterior zonules (white arrow) encroaching beyond the pupil margin in a dilated pupil. ( c ) Colour fundus photography shows areas of central atrophy affecting the macula and ( d ) Fundus autofluorescence imaging shows areas of central scalloped hypoautofluorescence matching the areas of atrophy surrounded by a cuff of increased autofluorescence. ( e ) Microperimetry interpolated plot superimposes areas of retinal sensitivity on a colour fundus image. Areas of good sensitvity vary from green to yellow to red for poor sensitivity. Areas with no sensitivity have no polygons. The image shows non-detectable sensitivity at the macula for the patient. ( f ) Hyperreflective deposits deep to the neural retina, indicated by asterisks, are observed in the macula in an OCT scan of the proband. NR, neural retina; CH, choroid. ( g ) Dark adaptometry of the proband (L-ORD case) shows that recovery of sensitivity after light exposure is slow and final sensitivity is still reduced at 80 minutes, compared to the normal, rapid recovery of sensitivity in an unaffected sibling. ( h ) Sequence analysis of C1QTNF5 exon 3 revealed a heterozygous c.646G > T transversion, resulting in a glycine to cysteine mutation at position 216 in the L-ORD case (lower panel) compared to an unaffected relative (upper panel). ( i ) HpyCh4V restriction digest confirms the c.646 G > T mutation by generating fragments of 439, 363, 129 and 120 bp in homozygous G (p.G216) samples, and 439, 363, 129, 94 and 26 bp for the mutant G > T (p.C216) allele.

    Techniques Used: Imaging, Sequencing, Mutagenesis

    6) Product Images from "SYBR Green Real-Time PCR-RFLP Assay Targeting the Plasmodium Cytochrome B Gene – A Highly Sensitive Molecular Tool for Malaria Parasite Detection and Species Determination"

    Article Title: SYBR Green Real-Time PCR-RFLP Assay Targeting the Plasmodium Cytochrome B Gene – A Highly Sensitive Molecular Tool for Malaria Parasite Detection and Species Determination

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0120210

    The cytb-qPCR products and RFLP assays for species determination. a) The 430 bp cytb-qPCR products for each species. b-e) Four RFLP assays using FspBI, AluI, HpyCH4V and Csp6I restriction enzymes for Plasmodium species determination. f) Schematic diagram showing the RFLP product lengths. a-e) Lane 1, GeneRuler low range DNA ladder (Thermo Fisher); F, P . falciparum ; V, P . vivax ; M, P . malariae ; O, P . ovale ; NC, negative control.
    Figure Legend Snippet: The cytb-qPCR products and RFLP assays for species determination. a) The 430 bp cytb-qPCR products for each species. b-e) Four RFLP assays using FspBI, AluI, HpyCH4V and Csp6I restriction enzymes for Plasmodium species determination. f) Schematic diagram showing the RFLP product lengths. a-e) Lane 1, GeneRuler low range DNA ladder (Thermo Fisher); F, P . falciparum ; V, P . vivax ; M, P . malariae ; O, P . ovale ; NC, negative control.

    Techniques Used: Real-time Polymerase Chain Reaction, Negative Control

    Related Articles

    other:

    Article Title: Restriction Analysis of β‐Tubulin Gene for Differentiation of the Common Pathogenic Dermatophytes
    Article Snippet: Likewise, in the current study, two species were nicely distinguished through BT2 fractionization by Hpy CH4V (Table , Fig. ).

    Polymerase Chain Reaction:

    Article Title: RDL mutations in Guangxi Anopheles sinensis populations along the China–Vietnam border: distribution frequency and evolutionary origin of A296S resistance allele
    Article Snippet: The reaction mixture (30 μl) included 2× Taq PCR MasterMix, 0.2 μM each primer and 30–100 ng of genomic DNA. .. The reaction procedure was set as 95 °C for 5 min; 38 cycles of 95 °C for 30 s, 56 °C for 30 s, 72 °C for 10 s; and 72 °C for 10 min. PCR products were digested with the restricted DNA digestion enzyme HpyCH4 III (New England Biolabs) (Fig. a) for 2 h in a total volume of 20 μl according to the manufacturer’s instruction. ..

    Article Title: Identification and distribution of a GABA receptor mutation conferring dieldrin resistance in the malaria vector Anopheles funestus in Africa
    Article Snippet: In order to provide a diagnostic assay that could be applied in laboratories without access to a pyrosequencer, we also designed a PCR-RFLP assay to genotype the A296S mutation. .. After amplification of the 255 bp fragment by PCR, the HpyCH4V restriction endonuclease cuts the susceptible allele to generate two fragments of 117 and 138 bp that co-migrate at the same position on a 2% agarose gel to give one band for G/G genotype ( ). ..

    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).

    Article Title: Identification and distribution of a GABA receptor mutation conferring dieldrin resistance in the malaria vector Anopheles funestus in Africa
    Article Snippet: The same primers RDLF and RDLR used above were used to amplify the fragment of 255 bp of Exon 7 using the same PCR conditions. .. The PCR product was directly digested with HpyCH4V restriction endonuclease (New England Biolabs) during 1 h of incubation at 37 °C. .. This restriction enzyme cleaves the susceptible allele at the recognition sequence TG^CA producing two fragments detectable on a 2% agarose gel.

    Amplification:

    Article Title: Rapid detection of equine coronavirus by reverse transcription loop-mediated isothermal amplification
    Article Snippet: .. To confirm the specificity of the RT-LAMP assays, amplified products were digested with HpyCH4V (New England Biolabs, Ipswich, MA, USA), a restriction enzyme. .. Digested products of expected lengths were observed using 2.2% (w/v) agarose gel electrophoresis (FlashGel System for DNA; Lonza Rockland) ( ).

    Article Title: Identification and distribution of a GABA receptor mutation conferring dieldrin resistance in the malaria vector Anopheles funestus in Africa
    Article Snippet: In order to provide a diagnostic assay that could be applied in laboratories without access to a pyrosequencer, we also designed a PCR-RFLP assay to genotype the A296S mutation. .. After amplification of the 255 bp fragment by PCR, the HpyCH4V restriction endonuclease cuts the susceptible allele to generate two fragments of 117 and 138 bp that co-migrate at the same position on a 2% agarose gel to give one band for G/G genotype ( ). ..

    Sequencing:

    Article Title: Seven novel mutations in the long isoform of the USH2A gene in Chinese families with nonsyndromic retinitis pigmentosa and Usher syndrome Type II
    Article Snippet: .. Restriction fragment length polymorphism analysis Variations (c.2802T > G, c.8232G > C, c.3788G > A, and c.14403C > G) found in the sequencing were confirmed with the restriction endonucleases Hinc II (TaKaRa, Dalian, China), HpyCH4V, BsaI, and SpeI (New England Biolabs, Ipswich, MA), respectively, which were used in all available family members and in the100 normal controls. .. Single strand conformation polymorphism To validate the variations (c. 1876C > T and c.7123delG) found in the sequencing, a single strand conformation polymorphism (SSCP) analysis was performed in all available family members and in the 100 normal controls.

    Agarose Gel Electrophoresis:

    Article Title: Identification and distribution of a GABA receptor mutation conferring dieldrin resistance in the malaria vector Anopheles funestus in Africa
    Article Snippet: In order to provide a diagnostic assay that could be applied in laboratories without access to a pyrosequencer, we also designed a PCR-RFLP assay to genotype the A296S mutation. .. After amplification of the 255 bp fragment by PCR, the HpyCH4V restriction endonuclease cuts the susceptible allele to generate two fragments of 117 and 138 bp that co-migrate at the same position on a 2% agarose gel to give one band for G/G genotype ( ). ..

    Incubation:

    Article Title: Identification and distribution of a GABA receptor mutation conferring dieldrin resistance in the malaria vector Anopheles funestus in Africa
    Article Snippet: The same primers RDLF and RDLR used above were used to amplify the fragment of 255 bp of Exon 7 using the same PCR conditions. .. The PCR product was directly digested with HpyCH4V restriction endonuclease (New England Biolabs) during 1 h of incubation at 37 °C. .. This restriction enzyme cleaves the susceptible allele at the recognition sequence TG^CA producing two fragments detectable on a 2% agarose gel.

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    New England Biolabs hpych4v restriction endonuclease
    Agarose gel of a PCR-RFLP to detect dieldrin resistance in An. funestus . The top band is a 255 bp fragment for resistant mosquitoes while the bottom bands represent the 117 and 138 bp fragments resulting from the restriction digestion by <t>HpyCH4V.</t>
    Hpych4v Restriction Endonuclease, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hpych4v restriction endonuclease/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hpych4v restriction endonuclease - by Bioz Stars, 2021-06
    95/100 stars
      Buy from Supplier

    Image Search Results


    Agarose gel of a PCR-RFLP to detect dieldrin resistance in An. funestus . The top band is a 255 bp fragment for resistant mosquitoes while the bottom bands represent the 117 and 138 bp fragments resulting from the restriction digestion by HpyCH4V.

    Journal: Insect Biochemistry and Molecular Biology

    Article Title: Identification and distribution of a GABA receptor mutation conferring dieldrin resistance in the malaria vector Anopheles funestus in Africa

    doi: 10.1016/j.ibmb.2011.03.012

    Figure Lengend Snippet: Agarose gel of a PCR-RFLP to detect dieldrin resistance in An. funestus . The top band is a 255 bp fragment for resistant mosquitoes while the bottom bands represent the 117 and 138 bp fragments resulting from the restriction digestion by HpyCH4V.

    Article Snippet: The PCR product was directly digested with HpyCH4V restriction endonuclease (New England Biolabs) during 1 h of incubation at 37 °C.

    Techniques: Agarose Gel Electrophoresis, Polymerase Chain Reaction

    Clonal Proliferation of HTLV-1-Infected Cells Can Be Detected by Agarose Gel Electrophoresis Genomic DNA was extracted from PBMCs from asymptomatic HTLV-1 carriers (n = 8), ATLL patients (n = 10), and a healthy individual, or for ATLL case 4 from a lymph node sample, and digested with HpyCH4V. Restriction fragments containing both the 3′ LTR of a provirus and human sequences were amplified as depicted in Figure 1 B and analyzed by agarose gel electrophoresis.

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: A Methodology for Assessing Tumor Clonality of Adult T Cell Leukemia/Lymphoma

    doi: 10.1016/j.omtm.2020.10.015

    Figure Lengend Snippet: Clonal Proliferation of HTLV-1-Infected Cells Can Be Detected by Agarose Gel Electrophoresis Genomic DNA was extracted from PBMCs from asymptomatic HTLV-1 carriers (n = 8), ATLL patients (n = 10), and a healthy individual, or for ATLL case 4 from a lymph node sample, and digested with HpyCH4V. Restriction fragments containing both the 3′ LTR of a provirus and human sequences were amplified as depicted in Figure 1 B and analyzed by agarose gel electrophoresis.

    Article Snippet: Genomic DNA (1 μg) was fragmented using the restriction enzyme HpyCH4V (New England Biolabs) at 37°C for 1 h. The enzyme was subsequently inactivated at 65°C for 20 min. NEBNext adaptors were ligated to the fragments using a NEBNext Ultra DNA Library Prep Kit for Illumina (New England Biolabs) and NEBNext Mulitiplex Oligos for Illumina (New England Biolabs).

    Techniques: Infection, Agarose Gel Electrophoresis, Amplification

    A restriction fragment length analysis of the four mutations detected in this study. A : c.2802T > G abolished a HincII restriction site that co-segregated with the affected individuals and the carriers (42 bp, 57 bp, 99 bp, 717 bp, and 774 bp), but not with unaffected individuals and normal controls (42 bp, 57 bp, and 717 bp). B : c.8232G > C created a new HpyCH4V restriction site that co-segregated with the affected individuals and the carriers (88 bp, 186 bp, 218 bp, and 274 bp), but not with unaffected individuals and normal controls (218 bp, 274 bp). C : c.3788G > A abolished a BsaI restriction site that co-segregated with the affected individuals and the carriers (70 bp, 132 bp, 422 bp, and 492 bp), but not with unaffected individuals and normal controls (70 bp, 132 bp, and 422 bp). D : c.14403C > G created a SpeI restriction site that co-segregated with the affected individuals and the carriers (145 bp, 300 bp, and 445 bp), but not with unaffected individuals and normal controls (445 bp). A participant identification number is given above each lane. N represents normal controls.

    Journal: Molecular Vision

    Article Title: Seven novel mutations in the long isoform of the USH2A gene in Chinese families with nonsyndromic retinitis pigmentosa and Usher syndrome Type II

    doi:

    Figure Lengend Snippet: A restriction fragment length analysis of the four mutations detected in this study. A : c.2802T > G abolished a HincII restriction site that co-segregated with the affected individuals and the carriers (42 bp, 57 bp, 99 bp, 717 bp, and 774 bp), but not with unaffected individuals and normal controls (42 bp, 57 bp, and 717 bp). B : c.8232G > C created a new HpyCH4V restriction site that co-segregated with the affected individuals and the carriers (88 bp, 186 bp, 218 bp, and 274 bp), but not with unaffected individuals and normal controls (218 bp, 274 bp). C : c.3788G > A abolished a BsaI restriction site that co-segregated with the affected individuals and the carriers (70 bp, 132 bp, 422 bp, and 492 bp), but not with unaffected individuals and normal controls (70 bp, 132 bp, and 422 bp). D : c.14403C > G created a SpeI restriction site that co-segregated with the affected individuals and the carriers (145 bp, 300 bp, and 445 bp), but not with unaffected individuals and normal controls (445 bp). A participant identification number is given above each lane. N represents normal controls.

    Article Snippet: Restriction fragment length polymorphism analysis Variations (c.2802T > G, c.8232G > C, c.3788G > A, and c.14403C > G) found in the sequencing were confirmed with the restriction endonucleases Hinc II (TaKaRa, Dalian, China), HpyCH4V, BsaI, and SpeI (New England Biolabs, Ipswich, MA), respectively, which were used in all available family members and in the100 normal controls.

    Techniques:

    Diagnostic PCR-RFLP tests to detect the T345S mutation. The 167 bp amplicon is undigested by HpyCH4 III for homozygous TCG (mutant 345SS), and cut into two fragments of 83 bp and 84 bp for homozygous ACG (wild 345TT) that co-migrate in the agarose gel and cannot be distinguished. A combined pattern (two bands 167 bp + 83/84 bp) is displayed for heterozygotes

    Journal: Malaria Journal

    Article Title: RDL mutations in Guangxi Anopheles sinensis populations along the China–Vietnam border: distribution frequency and evolutionary origin of A296S resistance allele

    doi: 10.1186/s12936-020-3098-y

    Figure Lengend Snippet: Diagnostic PCR-RFLP tests to detect the T345S mutation. The 167 bp amplicon is undigested by HpyCH4 III for homozygous TCG (mutant 345SS), and cut into two fragments of 83 bp and 84 bp for homozygous ACG (wild 345TT) that co-migrate in the agarose gel and cannot be distinguished. A combined pattern (two bands 167 bp + 83/84 bp) is displayed for heterozygotes

    Article Snippet: The reaction procedure was set as 95 °C for 5 min; 38 cycles of 95 °C for 30 s, 56 °C for 30 s, 72 °C for 10 s; and 72 °C for 10 min. PCR products were digested with the restricted DNA digestion enzyme HpyCH4 III (New England Biolabs) (Fig. a) for 2 h in a total volume of 20 μl according to the manufacturer’s instruction.

    Techniques: Diagnostic Assay, Polymerase Chain Reaction, Mutagenesis, Amplification, Agarose Gel Electrophoresis