tspri  (New England Biolabs)


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    Structured Review

    New England Biolabs tspri
    CD1a immunohistochemistry FFPE tissue section from LCH patients stained with CD1a. ( a ) Two samples that were V600E mutation (+) after both primary <t>PCR</t> amplification and <t>TspRI</t> followed by additional PCR amplification. Four of five samples in this category demonstrated up to 70 % tumor content. These cases were easy to detect by primary PCR. ( b ) Examples of staining of samples that were BRAF mutation (-) after primary PCR, but mutation (+) after TspRI treatment. Seven of eight such samples contained
    Tspri, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tspri/product/New England Biolabs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tspri - by Bioz Stars, 2022-05
    93/100 stars

    Images

    1) Product Images from "BRAF V600 mutations in Langerhans cell histiocytosis with a simple and unique assay"

    Article Title: BRAF V600 mutations in Langerhans cell histiocytosis with a simple and unique assay

    Journal: Diagnostic Pathology

    doi: 10.1186/s13000-016-0489-z

    CD1a immunohistochemistry FFPE tissue section from LCH patients stained with CD1a. ( a ) Two samples that were V600E mutation (+) after both primary PCR amplification and TspRI followed by additional PCR amplification. Four of five samples in this category demonstrated up to 70 % tumor content. These cases were easy to detect by primary PCR. ( b ) Examples of staining of samples that were BRAF mutation (-) after primary PCR, but mutation (+) after TspRI treatment. Seven of eight such samples contained
    Figure Legend Snippet: CD1a immunohistochemistry FFPE tissue section from LCH patients stained with CD1a. ( a ) Two samples that were V600E mutation (+) after both primary PCR amplification and TspRI followed by additional PCR amplification. Four of five samples in this category demonstrated up to 70 % tumor content. These cases were easy to detect by primary PCR. ( b ) Examples of staining of samples that were BRAF mutation (-) after primary PCR, but mutation (+) after TspRI treatment. Seven of eight such samples contained

    Techniques Used: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Staining, Mutagenesis, Polymerase Chain Reaction, Amplification

    The effect of TspRI restriction enzyme treatment on the BRAF sequence analysis of patient samples ( a ) Patient no. 21, representative sample with no V600E mutation, was judged mutation (-):GTG after primary PCR, and this conclusion was not changed by treatment with TspRI followed by secondary PCR; ( b ) patient no. 23 was scored as mutation (-):GTG after primary PCR, but mutation (+):GAG after treatment with TspRI; ( c ) the mutation status of patient no. 6 was difficult to judge from the primary PCR because of overlap between mutation (-):GTG and mutation (+):GAG sequences, however, after treatment with TspRI, the mutation (+):GAG sequence was dominant, indicating a clear mutation (+):GAG status; ( d ) patient no. 3 was initially judged mutation (-):GTG after primary PCR, but mutation (+):GAT status was apparent after treatment with TspRI
    Figure Legend Snippet: The effect of TspRI restriction enzyme treatment on the BRAF sequence analysis of patient samples ( a ) Patient no. 21, representative sample with no V600E mutation, was judged mutation (-):GTG after primary PCR, and this conclusion was not changed by treatment with TspRI followed by secondary PCR; ( b ) patient no. 23 was scored as mutation (-):GTG after primary PCR, but mutation (+):GAG after treatment with TspRI; ( c ) the mutation status of patient no. 6 was difficult to judge from the primary PCR because of overlap between mutation (-):GTG and mutation (+):GAG sequences, however, after treatment with TspRI, the mutation (+):GAG sequence was dominant, indicating a clear mutation (+):GAG status; ( d ) patient no. 3 was initially judged mutation (-):GTG after primary PCR, but mutation (+):GAT status was apparent after treatment with TspRI

    Techniques Used: Sequencing, Mutagenesis, Polymerase Chain Reaction

    Optimization of TspRI digestion for the BRAF amplicons. 100 ng of BRAF PCR fragments amplified from normal DNA were digested with TspRI. Lane 1: No TspRI, 65 °C, 15 min. Lane 2: 5U TspRI, 65 °C, 5 min. Lane 3: 10U TspRI, 65 °C, 10 min. Lane 4: 20U TspRI, 65 °C, 15 min, leading to digestion of the majority of BRAF PCR fragments
    Figure Legend Snippet: Optimization of TspRI digestion for the BRAF amplicons. 100 ng of BRAF PCR fragments amplified from normal DNA were digested with TspRI. Lane 1: No TspRI, 65 °C, 15 min. Lane 2: 5U TspRI, 65 °C, 5 min. Lane 3: 10U TspRI, 65 °C, 10 min. Lane 4: 20U TspRI, 65 °C, 15 min, leading to digestion of the majority of BRAF PCR fragments

    Techniques Used: Polymerase Chain Reaction, Amplification

    2) Product Images from "Efficient amplification of self-gelling polypod-like structured DNA by rolling circle amplification and enzymatic digestion"

    Article Title: Efficient amplification of self-gelling polypod-like structured DNA by rolling circle amplification and enzymatic digestion

    Journal: Scientific Reports

    doi: 10.1038/srep14979

    Schematic diagram of the mass amplification of tripodna with adhesive 5′-ends. The template oligodeoxynucleotides (template 1) were designed to satisfy the following requirements: ( a ) the polypodna automatically forms by self-assembly; ( b ) each pod of the polypodna contains a 9 base long TspRI restriction digest site; ( c ) Each 5′-terminal end is phosphorylated in order to ligate with 3′-terminal within the polypodna body, and ( d ) connecting chain is added to the 3′-terminal of the polypodna to allow polypodna to be connected to one another. The designed templates were amplified via the following steps. ( 1 ) The template ssODNs were circularized using T4 DNA ligase. ( 2 ) After annealing the primer (primer 1), the DNA template was amplified through rolling circle amplification technique using Phi29 polymerase. ( 3 ) Before enzyme digestion, the RCA product was treated with EDTA and folded. ( 4 ) Long single-stranded DNAs were digested using restriction enzyme. ( 5 ) The target sequences were purified by size chromatography. ( 6 ) The resultant DNAs self-assembled after annealing, and they formed a hydrogel.
    Figure Legend Snippet: Schematic diagram of the mass amplification of tripodna with adhesive 5′-ends. The template oligodeoxynucleotides (template 1) were designed to satisfy the following requirements: ( a ) the polypodna automatically forms by self-assembly; ( b ) each pod of the polypodna contains a 9 base long TspRI restriction digest site; ( c ) Each 5′-terminal end is phosphorylated in order to ligate with 3′-terminal within the polypodna body, and ( d ) connecting chain is added to the 3′-terminal of the polypodna to allow polypodna to be connected to one another. The designed templates were amplified via the following steps. ( 1 ) The template ssODNs were circularized using T4 DNA ligase. ( 2 ) After annealing the primer (primer 1), the DNA template was amplified through rolling circle amplification technique using Phi29 polymerase. ( 3 ) Before enzyme digestion, the RCA product was treated with EDTA and folded. ( 4 ) Long single-stranded DNAs were digested using restriction enzyme. ( 5 ) The target sequences were purified by size chromatography. ( 6 ) The resultant DNAs self-assembled after annealing, and they formed a hydrogel.

    Techniques Used: Amplification, Purification, Chromatography

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    New England Biolabs tspri
    CD1a immunohistochemistry FFPE tissue section from LCH patients stained with CD1a. ( a ) Two samples that were V600E mutation (+) after both primary <t>PCR</t> amplification and <t>TspRI</t> followed by additional PCR amplification. Four of five samples in this category demonstrated up to 70 % tumor content. These cases were easy to detect by primary PCR. ( b ) Examples of staining of samples that were BRAF mutation (-) after primary PCR, but mutation (+) after TspRI treatment. Seven of eight such samples contained
    Tspri, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tspri/product/New England Biolabs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    tspri - by Bioz Stars, 2022-05
    93/100 stars
      Buy from Supplier

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    CD1a immunohistochemistry FFPE tissue section from LCH patients stained with CD1a. ( a ) Two samples that were V600E mutation (+) after both primary PCR amplification and TspRI followed by additional PCR amplification. Four of five samples in this category demonstrated up to 70 % tumor content. These cases were easy to detect by primary PCR. ( b ) Examples of staining of samples that were BRAF mutation (-) after primary PCR, but mutation (+) after TspRI treatment. Seven of eight such samples contained

    Journal: Diagnostic Pathology

    Article Title: BRAF V600 mutations in Langerhans cell histiocytosis with a simple and unique assay

    doi: 10.1186/s13000-016-0489-z

    Figure Lengend Snippet: CD1a immunohistochemistry FFPE tissue section from LCH patients stained with CD1a. ( a ) Two samples that were V600E mutation (+) after both primary PCR amplification and TspRI followed by additional PCR amplification. Four of five samples in this category demonstrated up to 70 % tumor content. These cases were easy to detect by primary PCR. ( b ) Examples of staining of samples that were BRAF mutation (-) after primary PCR, but mutation (+) after TspRI treatment. Seven of eight such samples contained

    Article Snippet: Digestion of PCR products with TspRI was performed at 65 °C in 1 × CutSmart Buffer (New England Biolabs).

    Techniques: Immunohistochemistry, Formalin-fixed Paraffin-Embedded, Staining, Mutagenesis, Polymerase Chain Reaction, Amplification

    The effect of TspRI restriction enzyme treatment on the BRAF sequence analysis of patient samples ( a ) Patient no. 21, representative sample with no V600E mutation, was judged mutation (-):GTG after primary PCR, and this conclusion was not changed by treatment with TspRI followed by secondary PCR; ( b ) patient no. 23 was scored as mutation (-):GTG after primary PCR, but mutation (+):GAG after treatment with TspRI; ( c ) the mutation status of patient no. 6 was difficult to judge from the primary PCR because of overlap between mutation (-):GTG and mutation (+):GAG sequences, however, after treatment with TspRI, the mutation (+):GAG sequence was dominant, indicating a clear mutation (+):GAG status; ( d ) patient no. 3 was initially judged mutation (-):GTG after primary PCR, but mutation (+):GAT status was apparent after treatment with TspRI

    Journal: Diagnostic Pathology

    Article Title: BRAF V600 mutations in Langerhans cell histiocytosis with a simple and unique assay

    doi: 10.1186/s13000-016-0489-z

    Figure Lengend Snippet: The effect of TspRI restriction enzyme treatment on the BRAF sequence analysis of patient samples ( a ) Patient no. 21, representative sample with no V600E mutation, was judged mutation (-):GTG after primary PCR, and this conclusion was not changed by treatment with TspRI followed by secondary PCR; ( b ) patient no. 23 was scored as mutation (-):GTG after primary PCR, but mutation (+):GAG after treatment with TspRI; ( c ) the mutation status of patient no. 6 was difficult to judge from the primary PCR because of overlap between mutation (-):GTG and mutation (+):GAG sequences, however, after treatment with TspRI, the mutation (+):GAG sequence was dominant, indicating a clear mutation (+):GAG status; ( d ) patient no. 3 was initially judged mutation (-):GTG after primary PCR, but mutation (+):GAT status was apparent after treatment with TspRI

    Article Snippet: Digestion of PCR products with TspRI was performed at 65 °C in 1 × CutSmart Buffer (New England Biolabs).

    Techniques: Sequencing, Mutagenesis, Polymerase Chain Reaction

    Optimization of TspRI digestion for the BRAF amplicons. 100 ng of BRAF PCR fragments amplified from normal DNA were digested with TspRI. Lane 1: No TspRI, 65 °C, 15 min. Lane 2: 5U TspRI, 65 °C, 5 min. Lane 3: 10U TspRI, 65 °C, 10 min. Lane 4: 20U TspRI, 65 °C, 15 min, leading to digestion of the majority of BRAF PCR fragments

    Journal: Diagnostic Pathology

    Article Title: BRAF V600 mutations in Langerhans cell histiocytosis with a simple and unique assay

    doi: 10.1186/s13000-016-0489-z

    Figure Lengend Snippet: Optimization of TspRI digestion for the BRAF amplicons. 100 ng of BRAF PCR fragments amplified from normal DNA were digested with TspRI. Lane 1: No TspRI, 65 °C, 15 min. Lane 2: 5U TspRI, 65 °C, 5 min. Lane 3: 10U TspRI, 65 °C, 10 min. Lane 4: 20U TspRI, 65 °C, 15 min, leading to digestion of the majority of BRAF PCR fragments

    Article Snippet: Digestion of PCR products with TspRI was performed at 65 °C in 1 × CutSmart Buffer (New England Biolabs).

    Techniques: Polymerase Chain Reaction, Amplification

    Schematic diagram of the mass amplification of tripodna with adhesive 5′-ends. The template oligodeoxynucleotides (template 1) were designed to satisfy the following requirements: ( a ) the polypodna automatically forms by self-assembly; ( b ) each pod of the polypodna contains a 9 base long TspRI restriction digest site; ( c ) Each 5′-terminal end is phosphorylated in order to ligate with 3′-terminal within the polypodna body, and ( d ) connecting chain is added to the 3′-terminal of the polypodna to allow polypodna to be connected to one another. The designed templates were amplified via the following steps. ( 1 ) The template ssODNs were circularized using T4 DNA ligase. ( 2 ) After annealing the primer (primer 1), the DNA template was amplified through rolling circle amplification technique using Phi29 polymerase. ( 3 ) Before enzyme digestion, the RCA product was treated with EDTA and folded. ( 4 ) Long single-stranded DNAs were digested using restriction enzyme. ( 5 ) The target sequences were purified by size chromatography. ( 6 ) The resultant DNAs self-assembled after annealing, and they formed a hydrogel.

    Journal: Scientific Reports

    Article Title: Efficient amplification of self-gelling polypod-like structured DNA by rolling circle amplification and enzymatic digestion

    doi: 10.1038/srep14979

    Figure Lengend Snippet: Schematic diagram of the mass amplification of tripodna with adhesive 5′-ends. The template oligodeoxynucleotides (template 1) were designed to satisfy the following requirements: ( a ) the polypodna automatically forms by self-assembly; ( b ) each pod of the polypodna contains a 9 base long TspRI restriction digest site; ( c ) Each 5′-terminal end is phosphorylated in order to ligate with 3′-terminal within the polypodna body, and ( d ) connecting chain is added to the 3′-terminal of the polypodna to allow polypodna to be connected to one another. The designed templates were amplified via the following steps. ( 1 ) The template ssODNs were circularized using T4 DNA ligase. ( 2 ) After annealing the primer (primer 1), the DNA template was amplified through rolling circle amplification technique using Phi29 polymerase. ( 3 ) Before enzyme digestion, the RCA product was treated with EDTA and folded. ( 4 ) Long single-stranded DNAs were digested using restriction enzyme. ( 5 ) The target sequences were purified by size chromatography. ( 6 ) The resultant DNAs self-assembled after annealing, and they formed a hydrogel.

    Article Snippet: After purification by size-exclusion chromatography, the resultant large molecular weight DNA was digested with 0.1 U/μL TspRI (New England Biolabs) in solution containing 50 mM potassium acetate, 20 mM Tris-acetate, 10 mM magnesium acetate, and 100 μg/ml BSA.

    Techniques: Amplification, Purification, Chromatography