ecii  (New England Biolabs)


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  • 91
    Name:
    EciI
    Description:
    EciI 500 units
    Catalog Number:
    r0590l
    Price:
    269
    Size:
    500 units
    Category:
    Restriction Enzymes
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    Structured Review

    New England Biolabs ecii
    EciI
    EciI 500 units
    https://www.bioz.com/result/ecii/product/New England Biolabs
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    ecii - by Bioz Stars, 2020-07
    91/100 stars

    Images

    1) Product Images from "A Set of Assays for the Comprehensive Analysis of FMR1 Alleles in the Fragile X–Related Disorders"

    Article Title: A Set of Assays for the Comprehensive Analysis of FMR1 Alleles in the Fragile X–Related Disorders

    Journal: The Journal of Molecular Diagnostics : JMD

    doi: 10.1016/j.jmoldx.2016.06.001

    Analysis of the number of uninterrupted repeats at the 3′ end of the repeat tract using the AGG_RPT-PCR assay. A: Schematic representation of the basis of the AGG_RPT-PCR assay for AGG interruptions showing the FAM and HEX labeled strands generated by PCR that is the substrate for EciI digestion. B: Analysis of interruption status in A7306, a female carrier of a normal allele (30 repeats) and a gray zone allele (52 repeats) and HT-51E, a female carrier of a normal (29 repeats) and a PM allele (92 repeats). The products of the reaction were resolved by electrophoresis on a 2% agarose gel. C: Capillary electrophoretograms of the reaction products for HT-51E in which a FAM-labeled forward primer and a HEX-labeled reverse primer were used for the PCR. The CCG-rich strand of each fragment is shown in red and the CGG-rich strand is shown in blue. In the absence of EciI, the red and blue peaks correspond to the individual strands of the full-length PCR fragment. These fragments do not comigrate because the CGG strand has a higher mobility than the CCG strand in this gel system. After EciI digestion, the red peaks correspond to the CGG strand of the 3′ end of the repeat tract and the blue peaks correspond to the CCG strand of the 5′ end of the repeat tract. F, female; GZ, gray zone allele; MW, 100-bp molecular weight ladder; PM, premutation.
    Figure Legend Snippet: Analysis of the number of uninterrupted repeats at the 3′ end of the repeat tract using the AGG_RPT-PCR assay. A: Schematic representation of the basis of the AGG_RPT-PCR assay for AGG interruptions showing the FAM and HEX labeled strands generated by PCR that is the substrate for EciI digestion. B: Analysis of interruption status in A7306, a female carrier of a normal allele (30 repeats) and a gray zone allele (52 repeats) and HT-51E, a female carrier of a normal (29 repeats) and a PM allele (92 repeats). The products of the reaction were resolved by electrophoresis on a 2% agarose gel. C: Capillary electrophoretograms of the reaction products for HT-51E in which a FAM-labeled forward primer and a HEX-labeled reverse primer were used for the PCR. The CCG-rich strand of each fragment is shown in red and the CGG-rich strand is shown in blue. In the absence of EciI, the red and blue peaks correspond to the individual strands of the full-length PCR fragment. These fragments do not comigrate because the CGG strand has a higher mobility than the CCG strand in this gel system. After EciI digestion, the red peaks correspond to the CGG strand of the 3′ end of the repeat tract and the blue peaks correspond to the CCG strand of the 5′ end of the repeat tract. F, female; GZ, gray zone allele; MW, 100-bp molecular weight ladder; PM, premutation.

    Techniques Used: Polymerase Chain Reaction, Labeling, Generated, Electrophoresis, Agarose Gel Electrophoresis, Molecular Weight

    Related Articles

    Clone Assay:

    Article Title: Translation of the FMR1 mRNA is not influenced by AGG interruptions
    Article Snippet: .. The resulting plasmid constructs were confirmed both by sequencing and by EciI digestion (NEB) of the amplicon produced by PCR of the cloned region using standard FMR1 primers c and f ( ); EciI cleavage occurs 12 base pairs downstream of the A position in AGG interruptions. .. In addition to these three premutation-length, SP6-promoter FL plasmids, an analogous construct with 30 CGG repeats [designated pSP6-FMR1 (30CGG)-FL] was designed by XhoI, BlpI digestion of pSP6-FMR1 (65CGG)-FL, to remove the premutation-length repeat, followed by insertion of a similar digest containing 30 CGGs.

    Amplification:

    Article Title: Translation of the FMR1 mRNA is not influenced by AGG interruptions
    Article Snippet: .. The resulting plasmid constructs were confirmed both by sequencing and by EciI digestion (NEB) of the amplicon produced by PCR of the cloned region using standard FMR1 primers c and f ( ); EciI cleavage occurs 12 base pairs downstream of the A position in AGG interruptions. .. In addition to these three premutation-length, SP6-promoter FL plasmids, an analogous construct with 30 CGG repeats [designated pSP6-FMR1 (30CGG)-FL] was designed by XhoI, BlpI digestion of pSP6-FMR1 (65CGG)-FL, to remove the premutation-length repeat, followed by insertion of a similar digest containing 30 CGGs.

    Methylation:

    Article Title: Effect of aging on 5-hydroxymethylcytosine in the mouse hippocampus
    Article Snippet: .. The following enzymes were selected based on their published characteristics (REBASE Methylation Sensitivity: ) and their recognition sequences are indicated in parentheses: TseI [GCWGC], NmeAIII [GCCGAG (21/19)], EciI [GGCGGA (11/9)], SfaNI [GCATC (5/9)], and EcoP15I [CAGCAG (25/27)] (New England Biolabs; Ipswich, MA). .. Prior to restriction digestion, DNA was glucosylated with T4 glucosyl transferase (New England Biolabs) and aliquots were digested with the indicated enzymes in separate reactions.

    Construct:

    Article Title: Translation of the FMR1 mRNA is not influenced by AGG interruptions
    Article Snippet: .. The resulting plasmid constructs were confirmed both by sequencing and by EciI digestion (NEB) of the amplicon produced by PCR of the cloned region using standard FMR1 primers c and f ( ); EciI cleavage occurs 12 base pairs downstream of the A position in AGG interruptions. .. In addition to these three premutation-length, SP6-promoter FL plasmids, an analogous construct with 30 CGG repeats [designated pSP6-FMR1 (30CGG)-FL] was designed by XhoI, BlpI digestion of pSP6-FMR1 (65CGG)-FL, to remove the premutation-length repeat, followed by insertion of a similar digest containing 30 CGGs.

    Produced:

    Article Title: Translation of the FMR1 mRNA is not influenced by AGG interruptions
    Article Snippet: .. The resulting plasmid constructs were confirmed both by sequencing and by EciI digestion (NEB) of the amplicon produced by PCR of the cloned region using standard FMR1 primers c and f ( ); EciI cleavage occurs 12 base pairs downstream of the A position in AGG interruptions. .. In addition to these three premutation-length, SP6-promoter FL plasmids, an analogous construct with 30 CGG repeats [designated pSP6-FMR1 (30CGG)-FL] was designed by XhoI, BlpI digestion of pSP6-FMR1 (65CGG)-FL, to remove the premutation-length repeat, followed by insertion of a similar digest containing 30 CGGs.

    Sequencing:

    Article Title: Translation of the FMR1 mRNA is not influenced by AGG interruptions
    Article Snippet: .. The resulting plasmid constructs were confirmed both by sequencing and by EciI digestion (NEB) of the amplicon produced by PCR of the cloned region using standard FMR1 primers c and f ( ); EciI cleavage occurs 12 base pairs downstream of the A position in AGG interruptions. .. In addition to these three premutation-length, SP6-promoter FL plasmids, an analogous construct with 30 CGG repeats [designated pSP6-FMR1 (30CGG)-FL] was designed by XhoI, BlpI digestion of pSP6-FMR1 (65CGG)-FL, to remove the premutation-length repeat, followed by insertion of a similar digest containing 30 CGGs.

    Polymerase Chain Reaction:

    Article Title: Translation of the FMR1 mRNA is not influenced by AGG interruptions
    Article Snippet: .. The resulting plasmid constructs were confirmed both by sequencing and by EciI digestion (NEB) of the amplicon produced by PCR of the cloned region using standard FMR1 primers c and f ( ); EciI cleavage occurs 12 base pairs downstream of the A position in AGG interruptions. .. In addition to these three premutation-length, SP6-promoter FL plasmids, an analogous construct with 30 CGG repeats [designated pSP6-FMR1 (30CGG)-FL] was designed by XhoI, BlpI digestion of pSP6-FMR1 (65CGG)-FL, to remove the premutation-length repeat, followed by insertion of a similar digest containing 30 CGGs.

    Plasmid Preparation:

    Article Title: Translation of the FMR1 mRNA is not influenced by AGG interruptions
    Article Snippet: .. The resulting plasmid constructs were confirmed both by sequencing and by EciI digestion (NEB) of the amplicon produced by PCR of the cloned region using standard FMR1 primers c and f ( ); EciI cleavage occurs 12 base pairs downstream of the A position in AGG interruptions. .. In addition to these three premutation-length, SP6-promoter FL plasmids, an analogous construct with 30 CGG repeats [designated pSP6-FMR1 (30CGG)-FL] was designed by XhoI, BlpI digestion of pSP6-FMR1 (65CGG)-FL, to remove the premutation-length repeat, followed by insertion of a similar digest containing 30 CGGs.

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  • 91
    New England Biolabs ecii
    Analysis of the number of uninterrupted repeats at the 3′ end of the repeat tract using the <t>AGG_RPT-PCR</t> assay. A: Schematic representation of the basis of the AGG_RPT-PCR assay for AGG interruptions showing the FAM and HEX labeled strands generated by PCR that is the substrate for <t>EciI</t> digestion. B: Analysis of interruption status in A7306, a female carrier of a normal allele (30 repeats) and a gray zone allele (52 repeats) and HT-51E, a female carrier of a normal (29 repeats) and a PM allele (92 repeats). The products of the reaction were resolved by electrophoresis on a 2% agarose gel. C: Capillary electrophoretograms of the reaction products for HT-51E in which a FAM-labeled forward primer and a HEX-labeled reverse primer were used for the PCR. The CCG-rich strand of each fragment is shown in red and the CGG-rich strand is shown in blue. In the absence of EciI, the red and blue peaks correspond to the individual strands of the full-length PCR fragment. These fragments do not comigrate because the CGG strand has a higher mobility than the CCG strand in this gel system. After EciI digestion, the red peaks correspond to the CGG strand of the 3′ end of the repeat tract and the blue peaks correspond to the CCG strand of the 5′ end of the repeat tract. F, female; GZ, gray zone allele; MW, 100-bp molecular weight ladder; PM, premutation.
    Ecii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecii/product/New England Biolabs
    Average 91 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    ecii - by Bioz Stars, 2020-07
    91/100 stars
      Buy from Supplier

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    Analysis of the number of uninterrupted repeats at the 3′ end of the repeat tract using the AGG_RPT-PCR assay. A: Schematic representation of the basis of the AGG_RPT-PCR assay for AGG interruptions showing the FAM and HEX labeled strands generated by PCR that is the substrate for EciI digestion. B: Analysis of interruption status in A7306, a female carrier of a normal allele (30 repeats) and a gray zone allele (52 repeats) and HT-51E, a female carrier of a normal (29 repeats) and a PM allele (92 repeats). The products of the reaction were resolved by electrophoresis on a 2% agarose gel. C: Capillary electrophoretograms of the reaction products for HT-51E in which a FAM-labeled forward primer and a HEX-labeled reverse primer were used for the PCR. The CCG-rich strand of each fragment is shown in red and the CGG-rich strand is shown in blue. In the absence of EciI, the red and blue peaks correspond to the individual strands of the full-length PCR fragment. These fragments do not comigrate because the CGG strand has a higher mobility than the CCG strand in this gel system. After EciI digestion, the red peaks correspond to the CGG strand of the 3′ end of the repeat tract and the blue peaks correspond to the CCG strand of the 5′ end of the repeat tract. F, female; GZ, gray zone allele; MW, 100-bp molecular weight ladder; PM, premutation.

    Journal: The Journal of Molecular Diagnostics : JMD

    Article Title: A Set of Assays for the Comprehensive Analysis of FMR1 Alleles in the Fragile X–Related Disorders

    doi: 10.1016/j.jmoldx.2016.06.001

    Figure Lengend Snippet: Analysis of the number of uninterrupted repeats at the 3′ end of the repeat tract using the AGG_RPT-PCR assay. A: Schematic representation of the basis of the AGG_RPT-PCR assay for AGG interruptions showing the FAM and HEX labeled strands generated by PCR that is the substrate for EciI digestion. B: Analysis of interruption status in A7306, a female carrier of a normal allele (30 repeats) and a gray zone allele (52 repeats) and HT-51E, a female carrier of a normal (29 repeats) and a PM allele (92 repeats). The products of the reaction were resolved by electrophoresis on a 2% agarose gel. C: Capillary electrophoretograms of the reaction products for HT-51E in which a FAM-labeled forward primer and a HEX-labeled reverse primer were used for the PCR. The CCG-rich strand of each fragment is shown in red and the CGG-rich strand is shown in blue. In the absence of EciI, the red and blue peaks correspond to the individual strands of the full-length PCR fragment. These fragments do not comigrate because the CGG strand has a higher mobility than the CCG strand in this gel system. After EciI digestion, the red peaks correspond to the CGG strand of the 3′ end of the repeat tract and the blue peaks correspond to the CCG strand of the 5′ end of the repeat tract. F, female; GZ, gray zone allele; MW, 100-bp molecular weight ladder; PM, premutation.

    Article Snippet: A 10 μL aliquot of the standard RPT-PCR was digested with 2 U EciI or mock digested in a final volume of 20 μL containing 2 μL 10× CutSmart buffer (New England Biolabs).

    Techniques: Polymerase Chain Reaction, Labeling, Generated, Electrophoresis, Agarose Gel Electrophoresis, Molecular Weight