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
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Average 93 stars, based on 1 article reviews
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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
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
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
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
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