Restriction Enzyme Dde I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Impact of the Mitochondrial Genetic Background in Complex III Deficiency"
Article Title: Impact of the Mitochondrial Genetic Background in Complex III Deficiency
Journal: PLoS ONE
Figure Legend Snippet: Genetic and structural analysis of the m.15533A > G mutation. (A) Electropherogram showing the nucleotide change in patient's muscle DNA, indicated by arrowheads. (B) PCR-RFLP analysis of the MT-CYB mutation. Uncut (wild-type) DNA consists of a 150 bp PCR product. The mutated sequence contains one Dde I restriction site that results in two products of 90 and 60 bp after digestion. The control sequence contains one Mse I restriction site that results in the same two products after digestion. C, wild-type control; F, proband's fibroblasts DNA; M, proband's muscle DNA; Cy, control cybrid; 1, 2 and 3 refer to three independent mutant cybrid clones. (C) Alignment of cytochrome b amino acid sequences from selected species. Asparagine at amino acid position 263 is indicated with an arrow. (D) Partial 3D-images of the cytochrome b protein. The left panel shows the interaction site between the asparagine 263 (indicated in red) of cytochrome b and the aspartate 2 of the cytochrome c1 subunit (coloured in green). This is the closest negatively charged side-chain of cytochrome c1, which is facing the same aqueous pocket at a distance of 11.25 Armstrongs. Asparagine at position 260 of cytochrome b is indicated in blue. The central panel shows the 3D-structure of the asparagine 263 (in blue) in controls. The right panel shows the predicted structural effect of the N263D substitution (in red) in the patient. The images were obtained using the Chimera software  .
Techniques Used: Mutagenesis, Polymerase Chain Reaction, Sequencing, Software
Article Snippet: If a sufficient amount of DNA was observed, 8 μl of the PCR product was digested with the restriction enzyme Dde I (New England Biolabs) or doubly digested with
Article Snippet: After a pronounced band was obtained following electrophoresis, the PCR product was digested with 0.375 μl of the restriction enzyme Dde I (20,000 U/m; New England Biolabs, Inc., Beverly, Mass.) and analyzed by agarose gel electrophoresis with 4%
Article Snippet: To confirm that E. histolytica LAMP test amplified the predicted product, melt peaks were acquired using 1 °C steps, with a hold of 30 s, from 62 to 96 °C [ ] post amplification and through digestion of the resulting LAMP product using
Article Snippet: For the exon 3 c.140C > T (p.S47F) mutation, a region of 153 bp was amplified with primers FmU-S47F-F and FmU-S47F-R (Supplemental Table 1), and the amplicon was digested with the
Article Snippet: To quantify the heteroplasmy levels of the m.15533 A > G mutation, PCR-restriction fragment length polymorphism (RFLP) analysis was performed in the proband's family using the following primers: 5′-CACTATTCTCACCAGACCTC-3′ , (forward) and 5′-ACGCCTCCTAGTTTGTTAGG-3′ (reverse), and digestion with the