Noti Restriction Enzymes, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Analysis of Complex DNA Rearrangements During Early Stages of HAC Formation"
Article Title: Analysis of Complex DNA Rearrangements During Early Stages of HAC Formation
Figure Legend Snippet: Generation of synthetic α 21-I TetO and α 21-II LacO/Gal4 arrays. (A) Scheme of the pBAC11.32TW12.32GLII containing BAC and YAC cassettes, G418 resistance cassette and synthetic DNA: α 21-I TetO formed by high ordered repeats (HOR) monomers (green arrows) containing CENP-B boxes (blue) alternate with monomers containing TetO (yellow); α 21-II LacO/Gal4 formed by high ordered repeats (HOR) monomers (yellow arrows) containing Gal4 binding sequence (green) alternating with LacO (red). (B) Schematic of the assembly of the α 21-I TetO and α 21-II LacO/Gal4 arrays. (C, D) PFGE analysis of the nascent α 21-I TetO and α 21-II LacO/Gal4 arrays, cut with BamHI/NotI after each cycles of tandem ligation array amplification as described in Figure S2A (C) and Figure S2B (D). Expected sizes: α 21-I TetO 11-mer 1 copy (1.9 kb), 8 copies (15.2 kb), 32 copies (60.8 kb); α 21-II LacO/Gal4 12-mer 1 copy (2 kb), 8 copies (16 kb), 32 copies (64 kb). Plasmid vector is 2.9 kb, BAC vector is 7.1 kb. The asterisk (*) indicates the fragments that have been cloned into BAC vector (8 copies, 16 kb); red arrow in D indicates the size of the final pBAC11.32TW12.32GLII (∼120 kb) (m and M, markers).
Techniques Used: BAC Assay, Binding Assay, Sequencing, Ligation, Amplification, Plasmid Preparation, Clone Assay
Figure Legend Snippet: Screening of HT1080 colonies after transfection with pBAC11.32TW12.32GLII. (A) Scheme showing the possible fates of the pBAC11.32TW12.32GLII HAC seeding DNA after transfection in HT1080: in yellow and green (as integration or HAC) is represented the synthetic DNA. Timeline of the experiments performed from transfection into HT1080 cells. (B) BAC copy number (y axis) analyzed by qPCR in each HT1080 clone (x axis): only HT1080 clones containing > 20 BAC copies are represented in the graph. HT1080 clones are represented in green (HAC), red (integration) or mixture (both) according to the results of the FISH screening, as shown in C. Black arrows indicate the clones shown in C and analyzed further. (C) Representative pictures of oligo-FISH staining of HT1080 clones: slides have been hybridized with DNA probes (TetO-dig/rhodamine -dig antibody, Gal4-biotin and LacO-biotin/Fitc-streptavidin). DAPI stains DNA. Scalebar = 10µm. (D) Southern blot of selected HT1080 clonal DNA (as labelled on top of the panel) digested with BamHI and separated by CHEF; the transferred membrane was hybridized with radioactively labelled TetO (left) or LacO (right) specific probes. Red arrows indicate the expected size of the band without rearrangements. Clones labelled in red have been screened further (M and m, markers). (E) Cartoon of the pBAC11.32TW12.32GLII input DNA showing restriction sites for NotI and BamHI.
Techniques Used: Transfection, HAC Assay, BAC Assay, Real-time Polymerase Chain Reaction, Clone Assay, Fluorescence In Situ Hybridization, Staining, Southern Blot
Figure Legend Snippet: Formation of input pBAC11.32TW12.32GLII DNA. (A) CHEF analysis of 16 bacterial DNA after transformation with pBAC11.32TW12.32GLII and NotI and BamHI digestion: red arrows indicate the size of the final vector (∼120 kb); colonies labelled in red contain the insert of the desired length. DNA used for transfection as a control (in duplicate) (M marker). (B) Scheme of the pBAC11.32TW12.32GLII input DNA showing restriction sites for NotI and BamHI used to release the synthetic DNA. (C) PFGE analysis of selected bacterial colonies (in red) digested with EcoRI: each fragment originates from a different array (label on the left). DNA used for transfection as a control (in duplicate); original DNA as uncut sample (M marker). (D) α 21-I TetO and α 21-II LacO/Gal4 DNA ratio calculated with ImageJ on the intensity of the bands shown in C for each bacterial colony. Control and original DNA as in C. (E) CHEF analysis of bacterial colony #1 DNA (in duplicate) digested with NotI and BamHI to release the synthetic DNA (m and M, markers).
Techniques Used: Transformation Assay, Plasmid Preparation, Transfection, Marker
2) Product Images from "Transmission of Yersinia pseudotuberculosis in the Pork Production Chain from Farm to Slaughterhouse ▿"
Article Title: Transmission of Yersinia pseudotuberculosis in the Pork Production Chain from Farm to Slaughterhouse ▿
Journal: Applied and Environmental Microbiology
Figure Legend Snippet: Four different NotI profiles (lanes 1 to 4) of Yersinia pseudotuberculosis strains obtained. M designates midrange PFGE markers.
3) Product Images from "EphA2 targeting peptide tethered bioreducible poly(cystamine bisacrylamide - diamino hexane) for the delivery of therapeutic pCMV-RAE-1? to pancreatic islets"
Article Title: EphA2 targeting peptide tethered bioreducible poly(cystamine bisacrylamide - diamino hexane) for the delivery of therapeutic pCMV-RAE-1? to pancreatic islets
Journal: Journal of Controlled Release
Figure Legend Snippet: Digestion of pCMV-RAE-1γ with KpnI and NotI.