Structured Review

ATUM plasmid dna
Southern analysis of <t>transposon</t> insertions into S. cerevisiae genome. Genomic <t>DNA</t> of 17 G418-resistant yeast clones (diploid strain FY1679) was digested with BamHI + BglII (left) or HindIII (right) and probed with labeled kanMX4 DNA. (Lanes 1–17) Insertion mutants, (lane C) Genomic DNA of the original FY1679 recipient strain as a negative control, (lane P) Plasmid DNA containing the kanMX4-Mu transposon digested with HindIII as a positive control, fragment sizes on the left.
Plasmid Dna, supplied by ATUM, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/plasmid dna/product/ATUM
Average 86 stars, based on 1 article reviews
Price from $9.99 to $1999.99
plasmid dna - by Bioz Stars, 2021-03
86/100 stars

Images

1) Product Images from "Bacteriophage Mu integration in yeast and mammalian genomes"

Article Title: Bacteriophage Mu integration in yeast and mammalian genomes

Journal: Nucleic Acids Research

doi: 10.1093/nar/gkn801

Southern analysis of transposon insertions into S. cerevisiae genome. Genomic DNA of 17 G418-resistant yeast clones (diploid strain FY1679) was digested with BamHI + BglII (left) or HindIII (right) and probed with labeled kanMX4 DNA. (Lanes 1–17) Insertion mutants, (lane C) Genomic DNA of the original FY1679 recipient strain as a negative control, (lane P) Plasmid DNA containing the kanMX4-Mu transposon digested with HindIII as a positive control, fragment sizes on the left.
Figure Legend Snippet: Southern analysis of transposon insertions into S. cerevisiae genome. Genomic DNA of 17 G418-resistant yeast clones (diploid strain FY1679) was digested with BamHI + BglII (left) or HindIII (right) and probed with labeled kanMX4 DNA. (Lanes 1–17) Insertion mutants, (lane C) Genomic DNA of the original FY1679 recipient strain as a negative control, (lane P) Plasmid DNA containing the kanMX4-Mu transposon digested with HindIII as a positive control, fragment sizes on the left.

Techniques Used: Clone Assay, Labeling, Negative Control, Plasmid Preparation, Positive Control

Analysis of transpososome-mediated gene delivery into human ES cells. ( A ) Expression of eGFP. Human ES cell line FES29 was electroporated with Puro-eGFP-Mu transposons and selected for 2 days with puromycin. Surviving fluorescent colonies were isolated and further cultured as clonal cell lines for several passages. Most of the colonies of the clonal isolates showed uniform GFP expression. Two example clones are shown in the phase contrast (left) and fluorescent (right) micrographs. ( B ) Southern analysis of the insertions into the hES cell genome. Genomic DNA of nine puromycin-resistant hES cell clones was digested with BglII (left) or EcoRI (right) and probed with labeled Puro-eGFP-Mu transposon DNA. (Lane P) Undigested genomic DNA of clone 2 spiked with transposon DNA as a positive control. (M) Size marker.
Figure Legend Snippet: Analysis of transpososome-mediated gene delivery into human ES cells. ( A ) Expression of eGFP. Human ES cell line FES29 was electroporated with Puro-eGFP-Mu transposons and selected for 2 days with puromycin. Surviving fluorescent colonies were isolated and further cultured as clonal cell lines for several passages. Most of the colonies of the clonal isolates showed uniform GFP expression. Two example clones are shown in the phase contrast (left) and fluorescent (right) micrographs. ( B ) Southern analysis of the insertions into the hES cell genome. Genomic DNA of nine puromycin-resistant hES cell clones was digested with BglII (left) or EcoRI (right) and probed with labeled Puro-eGFP-Mu transposon DNA. (Lane P) Undigested genomic DNA of clone 2 spiked with transposon DNA as a positive control. (M) Size marker.

Techniques Used: Expressing, Isolation, Cell Culture, Clone Assay, Labeling, Positive Control, Marker

Southern blot analysis of transposon insertions into HeLa cell genome. Chromosomal DNA was doubly digested with BamHI + BglII and probed with labeled Kan/Neo-p15A-Mu transposon DNA. A total of 19 different G418-resistant clones are analyzed, some with their siblings (bracketed) for the verification of clonality. (Lane C) Genomic DNA of the recipient HeLa cell line as a negative control. (Lane P) HeLa cell DNA spiked with transposon DNA as a positive control. (Lane M) Size marker.
Figure Legend Snippet: Southern blot analysis of transposon insertions into HeLa cell genome. Chromosomal DNA was doubly digested with BamHI + BglII and probed with labeled Kan/Neo-p15A-Mu transposon DNA. A total of 19 different G418-resistant clones are analyzed, some with their siblings (bracketed) for the verification of clonality. (Lane C) Genomic DNA of the recipient HeLa cell line as a negative control. (Lane P) HeLa cell DNA spiked with transposon DNA as a positive control. (Lane M) Size marker.

Techniques Used: Southern Blot, Labeling, Clone Assay, Negative Control, Positive Control, Marker

Analysis of transpososome-mediated gene delivery into mouse ES cells. ( A ) Efficiency. Pre-assembled transpososomes made with Kan/Neo-p15A-Mu transposon were introduced into mouse cells by electroporation. Following G418 selection, surviving cell colonies were stained with methylene blue. Gene delivery was analyzed using transpososomes made with wild-type MuA protein (left) and active site mutant MuA(E392Q) (middle). Analysis was done also with linear transposon DNA (right). ( B ) Southern analysis of transposon insertions into the mES cell genome. Genomic DNA of 17 G418-resistant mES cell clones was doubly digested with BamHI + BglII and probed with labeled Kan/Neo-p15A-Mu transposon DNA. (Lanes 1–17) Transposon insertion mutants. (Lane C) Genomic DNA of the original AB2.2 recipient strain as a negative control. (Lane P) AB2.2 genomic DNA spiked with transposon DNA as a positive control. (Lane M) Size marker. The cross-hybridizing band present in all genomic DNA samples served as a loading control.
Figure Legend Snippet: Analysis of transpososome-mediated gene delivery into mouse ES cells. ( A ) Efficiency. Pre-assembled transpososomes made with Kan/Neo-p15A-Mu transposon were introduced into mouse cells by electroporation. Following G418 selection, surviving cell colonies were stained with methylene blue. Gene delivery was analyzed using transpososomes made with wild-type MuA protein (left) and active site mutant MuA(E392Q) (middle). Analysis was done also with linear transposon DNA (right). ( B ) Southern analysis of transposon insertions into the mES cell genome. Genomic DNA of 17 G418-resistant mES cell clones was doubly digested with BamHI + BglII and probed with labeled Kan/Neo-p15A-Mu transposon DNA. (Lanes 1–17) Transposon insertion mutants. (Lane C) Genomic DNA of the original AB2.2 recipient strain as a negative control. (Lane P) AB2.2 genomic DNA spiked with transposon DNA as a positive control. (Lane M) Size marker. The cross-hybridizing band present in all genomic DNA samples served as a loading control.

Techniques Used: Electroporation, Selection, Staining, Mutagenesis, Clone Assay, Labeling, Negative Control, Positive Control, Marker

2) Product Images from "Endocytosis of DNA-Hsp65 Alters the pH of the Late Endosome/Lysosome and Interferes with Antigen Presentation"

Article Title: Endocytosis of DNA-Hsp65 Alters the pH of the Late Endosome/Lysosome and Interferes with Antigen Presentation

Journal: PLoS ONE

doi: 10.1371/journal.pone.0000923

DNA treatment impairs KLH antigen presentation. Peritoneal macrophages were incubated with DNA (20 µg) 72 h, 48 h, 24 h or 0 h prior to the treatment with KLH (100 µg). After KLH treatment (24 h), the peritoneal macrophages were fixed and CD4 T cells specific for KLH were added to the culture. Proliferation was measured after 72 h. Concanavalin A (40 µg/ml) was used as positive control. * p
Figure Legend Snippet: DNA treatment impairs KLH antigen presentation. Peritoneal macrophages were incubated with DNA (20 µg) 72 h, 48 h, 24 h or 0 h prior to the treatment with KLH (100 µg). After KLH treatment (24 h), the peritoneal macrophages were fixed and CD4 T cells specific for KLH were added to the culture. Proliferation was measured after 72 h. Concanavalin A (40 µg/ml) was used as positive control. * p

Techniques Used: Incubation, Positive Control

3) Product Images from "Antifibrotic Effect of MMP13-encoding Plasmid DNA Delivered Using Polyethylenimine Shielded With Hyaluronic Acid"

Article Title: Antifibrotic Effect of MMP13-encoding Plasmid DNA Delivered Using Polyethylenimine Shielded With Hyaluronic Acid

Journal: Molecular Therapy

doi: 10.1038/mt.2010.262

Liver distribution of pMMP13 in PEI or HA/PEI complexes . Mice were intravenously injected with pMMP13 (1 mg/kg) in PEI complexes or HA-shielded PEI complexes. At 4 hours after injection, gDNA was extracted from blood and liver tissues. pMMP13 copy numbers in the liver were divided by those in the blood. The data are the mean ± SE ( n = 5). gDNA, genomic DNA; HA, hyaluronic acid; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.
Figure Legend Snippet: Liver distribution of pMMP13 in PEI or HA/PEI complexes . Mice were intravenously injected with pMMP13 (1 mg/kg) in PEI complexes or HA-shielded PEI complexes. At 4 hours after injection, gDNA was extracted from blood and liver tissues. pMMP13 copy numbers in the liver were divided by those in the blood. The data are the mean ± SE ( n = 5). gDNA, genomic DNA; HA, hyaluronic acid; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.

Techniques Used: Mouse Assay, Injection, Plasmid Preparation

Survival rates of mice after intravenous administration of pMMP13 in PEI or HA/PEI complexes . Mice in each group ( n = 5) were intravenously injected with pMMP13 in PEI or HA-shielded PEI complexes. The behavior changes of survived mice were checked for 1-month postdose. HA, hyaluronic acid; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.
Figure Legend Snippet: Survival rates of mice after intravenous administration of pMMP13 in PEI or HA/PEI complexes . Mice in each group ( n = 5) were intravenously injected with pMMP13 in PEI or HA-shielded PEI complexes. The behavior changes of survived mice were checked for 1-month postdose. HA, hyaluronic acid; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.

Techniques Used: Mouse Assay, Injection, Plasmid Preparation

Expression of MMP13 proteins in liver tissues of fibrotic mice . Expression of EGFP and MMP13 proteins in liver tissues was visualized by fluorescent microscopy and immunoblotting, respectively. ( a , c ) Representative phase-contrast and ( b , d ) fluorescence images were shown for tissues from ( a , b ) untreated liver or ( c , d ) pMMP13-treated liver. Bar = 100 µm. The expression of MMP13 proteins in liver tissues was analyzed by ( e ) immunoblotting. EGFP, enriched green fluorescent protein; HA, hyaluronic acid; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.
Figure Legend Snippet: Expression of MMP13 proteins in liver tissues of fibrotic mice . Expression of EGFP and MMP13 proteins in liver tissues was visualized by fluorescent microscopy and immunoblotting, respectively. ( a , c ) Representative phase-contrast and ( b , d ) fluorescence images were shown for tissues from ( a , b ) untreated liver or ( c , d ) pMMP13-treated liver. Bar = 100 µm. The expression of MMP13 proteins in liver tissues was analyzed by ( e ) immunoblotting. EGFP, enriched green fluorescent protein; HA, hyaluronic acid; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.

Techniques Used: Expressing, Mouse Assay, Microscopy, Fluorescence, Plasmid Preparation

Stability of pMMP13 in vitro and in vivo . ( a ) Naked pMMP13, PEI/pMMP13, or HA/PEI/pMMP13 was incubated with DNase I. The samples were collected at various time points. pMMP13 was extracted from the samples and loaded onto a 1% agarose gel. ( b ) Mice were injected intravenously with pMMP13 at 1 mg/kg dose in naked form or complexes. At 4 hours after injection, DNA was isolated from blood and pMMP13 copy numbers were determined by quantitative RT-PCR. The data are the mean ± SE ( n = 5). *Significantly higher than naked pMMP13 ( P
Figure Legend Snippet: Stability of pMMP13 in vitro and in vivo . ( a ) Naked pMMP13, PEI/pMMP13, or HA/PEI/pMMP13 was incubated with DNase I. The samples were collected at various time points. pMMP13 was extracted from the samples and loaded onto a 1% agarose gel. ( b ) Mice were injected intravenously with pMMP13 at 1 mg/kg dose in naked form or complexes. At 4 hours after injection, DNA was isolated from blood and pMMP13 copy numbers were determined by quantitative RT-PCR. The data are the mean ± SE ( n = 5). *Significantly higher than naked pMMP13 ( P

Techniques Used: In Vitro, In Vivo, Incubation, Agarose Gel Electrophoresis, Mouse Assay, Injection, Isolation, Quantitative RT-PCR

Schematic representation for the construction of pMMP13 and formation of HA-shielded PEI and plasmid DNA ternary complex for gene delivery . ( a ) MMP13 cDNA was inserted into the Bgl II/ Sal I sites of the pIRES2-EGFP expression vector encoding EGFP. The recombinant plasmid DNA encoding both MMP13 and EGFP was abbreviated as pMMP13. ( b ) Plasmid DNA was electrostatically complexed to cationic PEI, resulting in binary complexes. The binary complexes with surplus cationic charges were then coated with negatively charged HA, providing ternary complexes. cDNA, complementary DNA; EGFP, enriched green fluorescent protein; HA, hyaluronic acid; IRES, internal ribosome entry site; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.
Figure Legend Snippet: Schematic representation for the construction of pMMP13 and formation of HA-shielded PEI and plasmid DNA ternary complex for gene delivery . ( a ) MMP13 cDNA was inserted into the Bgl II/ Sal I sites of the pIRES2-EGFP expression vector encoding EGFP. The recombinant plasmid DNA encoding both MMP13 and EGFP was abbreviated as pMMP13. ( b ) Plasmid DNA was electrostatically complexed to cationic PEI, resulting in binary complexes. The binary complexes with surplus cationic charges were then coated with negatively charged HA, providing ternary complexes. cDNA, complementary DNA; EGFP, enriched green fluorescent protein; HA, hyaluronic acid; IRES, internal ribosome entry site; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.

Techniques Used: Plasmid Preparation, Expressing, Recombinant

Levels of MMP13 mRNA in liver tissues of fibrotic mice . ( a ) For induction of fibrosis, mice were intraperitoneally injected with CCl 4 and intravenously treated with HA/PEI/pVector or HA/PEI/pMMP13 according to the dosing schedule. The mRNA levels of MMP13 were measured from total RNA isolated from the liver tissue from normal mice, untreated fibrotic mice, or fibrotic mice treated with HA/PEI/pVector or HA/PEI/pMMP13. ( b ) The expression levels of MMP13 mRNA were normalized to those of GAPDH. The data are the mean ± SE ( n = 4). HA, hyaluronic acid; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.
Figure Legend Snippet: Levels of MMP13 mRNA in liver tissues of fibrotic mice . ( a ) For induction of fibrosis, mice were intraperitoneally injected with CCl 4 and intravenously treated with HA/PEI/pVector or HA/PEI/pMMP13 according to the dosing schedule. The mRNA levels of MMP13 were measured from total RNA isolated from the liver tissue from normal mice, untreated fibrotic mice, or fibrotic mice treated with HA/PEI/pVector or HA/PEI/pMMP13. ( b ) The expression levels of MMP13 mRNA were normalized to those of GAPDH. The data are the mean ± SE ( n = 4). HA, hyaluronic acid; PEI, polyethylenimine; pMMP13, plasmid DNA encoding matrix metalloproteinases 13.

Techniques Used: Mouse Assay, Injection, Isolation, Expressing, Plasmid Preparation

4) Product Images from "A practical device for pinpoint delivery of molecules into multiple neurons in culture"

Article Title: A practical device for pinpoint delivery of molecules into multiple neurons in culture

Journal: Brain Cell Biology

doi: 10.1007/s11068-008-9021-z

Delivery of chemicals, proteins, or DNA into HeLa cells. (A, B) Delivery of Rhodamine101. A PC image of a confluent monolayer of HeLa cells. Stab points are indicated by red crosses (A). A fluorescence image acquired immediately after washing the medium (B). (C, D) Delivery of recombinant DsRed. A PC image of a colony of HeLa cells. Stabbed points (cytosol) are indicated by red crosses (C). A fluorescence image acquired immediately after washing the medium (D). (E, F) A DIC image (E) and fluorescence image (F) of a colony of HeLa cells 24 h post-transfection with Venus plasmid DNA. (G, H) A DIC image (G) and fluorescence image (H) of HeLa cells 24 h after differential transfection of Venus and mRFP1 plasmid DNA. Scale bar, 50 μm
Figure Legend Snippet: Delivery of chemicals, proteins, or DNA into HeLa cells. (A, B) Delivery of Rhodamine101. A PC image of a confluent monolayer of HeLa cells. Stab points are indicated by red crosses (A). A fluorescence image acquired immediately after washing the medium (B). (C, D) Delivery of recombinant DsRed. A PC image of a colony of HeLa cells. Stabbed points (cytosol) are indicated by red crosses (C). A fluorescence image acquired immediately after washing the medium (D). (E, F) A DIC image (E) and fluorescence image (F) of a colony of HeLa cells 24 h post-transfection with Venus plasmid DNA. (G, H) A DIC image (G) and fluorescence image (H) of HeLa cells 24 h after differential transfection of Venus and mRFP1 plasmid DNA. Scale bar, 50 μm

Techniques Used: Fluorescence, Recombinant, Transfection, Plasmid Preparation

5) Product Images from "Inducible model for ?-six-mediated site-specific recombination in mammalian cells"

Article Title: Inducible model for ?-six-mediated site-specific recombination in mammalian cells

Journal: Nucleic Acids Research

doi: 10.1093/nar/gnj001

Analysis of recombination time-course. Several C10 subclones were obtained and their recombination rates analyzed over a 7-week period. The recombination rate obtained for five of them (10.2, 10.3, 10.5, 10.6 and 10.7) was analyzed by several techniques. ( A ) Non-recombined (NR) and recombined (R) DNA fragments amplified by PCR. ( B ) Detection of β recombinase expression and histone H1, as control, by western blot. ( C ) Luciferase activity. Western blot and luciferase activity were measured in the fifth week.
Figure Legend Snippet: Analysis of recombination time-course. Several C10 subclones were obtained and their recombination rates analyzed over a 7-week period. The recombination rate obtained for five of them (10.2, 10.3, 10.5, 10.6 and 10.7) was analyzed by several techniques. ( A ) Non-recombined (NR) and recombined (R) DNA fragments amplified by PCR. ( B ) Detection of β recombinase expression and histone H1, as control, by western blot. ( C ) Luciferase activity. Western blot and luciferase activity were measured in the fifth week.

Techniques Used: Amplification, Polymerase Chain Reaction, Expressing, Western Blot, Luciferase, Activity Assay

Analysis of recombination in psps-Luc-transduced NIH-3T3 clones. Ten clones (1–10) were isolated and SSR analyzed by PCR and luciferase expression. ( A ) Genomic DNA of each clone (−, untransduced cells; +, transduced cells) was amplified using specific primers of the EF-1α promoter and luciferase gene (see Figure 2A ). Non-recombined DNA (NR; 1.5 kb band), recombined DNA (R; 0.4 kb band) (upper panel). The recombination level was determined by luciferase activity quantification (lower panel). ( B ) EGFP-positive subpopulations were purified by cell sorting of three different clones (7, 8 and 10). Recombination level in untransduced (−) and transduced (pre-sorting and post-sorting) cells was analyzed by PCR.
Figure Legend Snippet: Analysis of recombination in psps-Luc-transduced NIH-3T3 clones. Ten clones (1–10) were isolated and SSR analyzed by PCR and luciferase expression. ( A ) Genomic DNA of each clone (−, untransduced cells; +, transduced cells) was amplified using specific primers of the EF-1α promoter and luciferase gene (see Figure 2A ). Non-recombined DNA (NR; 1.5 kb band), recombined DNA (R; 0.4 kb band) (upper panel). The recombination level was determined by luciferase activity quantification (lower panel). ( B ) EGFP-positive subpopulations were purified by cell sorting of three different clones (7, 8 and 10). Recombination level in untransduced (−) and transduced (pre-sorting and post-sorting) cells was analyzed by PCR.

Techniques Used: Clone Assay, Isolation, Polymerase Chain Reaction, Luciferase, Expressing, Amplification, Activity Assay, Purification, FACS

Related Articles

Transfection:

Article Title: Four key steps control glycolytic flux in mammalian cells
Article Snippet: Correspondingly, serial dilutions of a given construct were supplemented with vector plasmid DNA to keep the 2.5 μg total plasmid DNA amount. iBMK cells were incubated with the plasmid DNA-lipofectamine complexes in 2 ml DMEM (10% dFBS) for 6 h before switching to fresh DMEM (10% dFBS). .. For transfection in 6 cm dishes, 1 μl PCV iBMK cells were plated and grown overnight before transfection with 8 μg of plasmid DNA, 20 μl of lipofectamine as well as 500 μl of OptiMEM incubated in 5 ml of DMEM (10% dFBS) for 6 h. Number of viable cells was counted at the indicated time points. .. Cells were transfected in 6-well plates as described above and viable cells were counted using Trypan blue exclusion assays and a Countess™ II FL Automated Cell Counter (ThermoFisher, MA).

Article Title: Assembly and Folding Properties of Cytosolic IgG Intrabodies
Article Snippet: After 6–7 days, the culture supernatant was harvested by centrifugation, and Ig proteins were purified by affinity chromatography using agarose-Protein A (GE Healthcare; cat# 17-1279-02) according to the manufacturer’s instructions. .. Analysis of the redox state of IgGs HEK293 cells, seeded in 100 mm dishes at a density of 5 × 106 cells/dish, were transfected by incubating with a mixture of plasmid DNA (20 μg) plus PEI reagent (96 μg) in 10 ml Opti-MEM at 37 °C for 24 hours. .. Cells were pre-incubated with NEM at a final concentration of 50 mM at 37 °C for 15 min and lysed by sonication as described above in ‘Preparation of cell lysates ’ in the presence of 100 mM NEM.

Plasmid Preparation:

Article Title: Four key steps control glycolytic flux in mammalian cells
Article Snippet: Correspondingly, serial dilutions of a given construct were supplemented with vector plasmid DNA to keep the 2.5 μg total plasmid DNA amount. iBMK cells were incubated with the plasmid DNA-lipofectamine complexes in 2 ml DMEM (10% dFBS) for 6 h before switching to fresh DMEM (10% dFBS). .. For transfection in 6 cm dishes, 1 μl PCV iBMK cells were plated and grown overnight before transfection with 8 μg of plasmid DNA, 20 μl of lipofectamine as well as 500 μl of OptiMEM incubated in 5 ml of DMEM (10% dFBS) for 6 h. Number of viable cells was counted at the indicated time points. .. Cells were transfected in 6-well plates as described above and viable cells were counted using Trypan blue exclusion assays and a Countess™ II FL Automated Cell Counter (ThermoFisher, MA).

Article Title: The architecture of the human Rad54-DNA complex provides evidence for protein translocation along DNA
Article Snippet: Plasmid pDERI1 was generated by deletion of the Ssp I– Sap I fragment from plasmid pUC19, resulting in a plasmid 1,821 bp in length. .. Singly nicked plasmid DNA was produced in a 30-μl reaction mixture containing 0.5 μg of DNA, 20 mM Tris⋅HCl (pH 7.5), 50 mM NaCl, 10 mM MgCl2 , 360 μg/ml ethidium bromide, and 1 μg/ml DNase I at 30°C for 30 min. .. The reaction was stopped by the addition of 0.1 vol of 5% (wt/vol) SDS/50 mM EDTA/30 μg/ml proteinase K and subsequent incubation at 65°C for 30 min. DNA was purified by extraction with phenol and phenol/chloroform (1:1, vol/vol), precipitated with ethanol, and dissolved in 10 mM Tris⋅HCl (pH 8.0)/1 mM EDTA.

Article Title: MicroRNA miR-29 controls a compensatory response to limit neuronal iron accumulation during adult life and aging
Article Snippet: Capillaries were pulled with a micropipette puller (P97, Sutter Instrument). .. The needles were filled with 2μl of water solution containing 20-30 ng/μl of plasmid DNA, 20-30 ng/μl of Tol2 transposase mRNA, 0,4M KCI and 1% phenol red as visual control of successful injections. .. The embryos were injected with ~2nL of mix solution and the drop volume was estimated under a microscope using a calibrated slide.).

Article Title: Synthetic biology based construction of biological activity-related library of fungal decalin-containing diterpenoid pyrones
Article Snippet: .. 0.2 Volumes of solution II (40% PEG4000, 50 mM CaCl2 , 50 mM Tris-HCl (pH 8.0)) was added with gentle mixing, and then 200 µL of this protoplast suspension was combined with 10–20 µg of plasmid DNA ( < 20 µL). .. After 40 min incubation on ice, 1 mL of solution II was added to the aliquot with gentle mixing.

Article Title: Bacteriophage Mu integration in yeast and mammalian genomes
Article Snippet: .. An aliquot (1–2 μl) of transpososome preparation (containing 1 μg transposon DNA) or plasmid DNA (20 ng) was added to the cell suspension. .. Following incubation on ice (5 min), the mixture was transferred into a chilled Bio-Rad cuvette (0.2-cm electrode spacing), and electroporation was carried out using Genepulser II (Bio-Rad, Hercules, CA, USA) with the following settings: voltage 1.5 kV (diploid strain FY1679) or 2.0 kV (haploid strain FY-3a); capacitance 25 μF; resistance 200 Ω.

Article Title: DNA Methyltransferase 3a and Mitogen-activated Protein Kinase Signaling Regulate the Expression of Fibroblast Growth Factor-inducible 14 (Fn14) during Denervation-induced Skeletal Muscle Atrophy *
Article Snippet: In brief, plasmids were prepared using an endotoxin-free kit (Qiagen) and suspended in sterile saline solution. .. The mice were anesthetized, and a small portion of TA muscle of both hind limbs was exposed and injected with plasmid DNA (20 μg in 20 μl of saline). .. After 1 min of plasmid DNA injection, a pair of platinum plate electrodes was placed against the closely shaved skin on either side of the small surgical incision, and electric pulses were delivered transcutaneously.

Article Title: Assembly and Folding Properties of Cytosolic IgG Intrabodies
Article Snippet: After 6–7 days, the culture supernatant was harvested by centrifugation, and Ig proteins were purified by affinity chromatography using agarose-Protein A (GE Healthcare; cat# 17-1279-02) according to the manufacturer’s instructions. .. Analysis of the redox state of IgGs HEK293 cells, seeded in 100 mm dishes at a density of 5 × 106 cells/dish, were transfected by incubating with a mixture of plasmid DNA (20 μg) plus PEI reagent (96 μg) in 10 ml Opti-MEM at 37 °C for 24 hours. .. Cells were pre-incubated with NEM at a final concentration of 50 mM at 37 °C for 15 min and lysed by sonication as described above in ‘Preparation of cell lysates ’ in the presence of 100 mM NEM.

Incubation:

Article Title: Four key steps control glycolytic flux in mammalian cells
Article Snippet: Correspondingly, serial dilutions of a given construct were supplemented with vector plasmid DNA to keep the 2.5 μg total plasmid DNA amount. iBMK cells were incubated with the plasmid DNA-lipofectamine complexes in 2 ml DMEM (10% dFBS) for 6 h before switching to fresh DMEM (10% dFBS). .. For transfection in 6 cm dishes, 1 μl PCV iBMK cells were plated and grown overnight before transfection with 8 μg of plasmid DNA, 20 μl of lipofectamine as well as 500 μl of OptiMEM incubated in 5 ml of DMEM (10% dFBS) for 6 h. Number of viable cells was counted at the indicated time points. .. Cells were transfected in 6-well plates as described above and viable cells were counted using Trypan blue exclusion assays and a Countess™ II FL Automated Cell Counter (ThermoFisher, MA).

Produced:

Article Title: The architecture of the human Rad54-DNA complex provides evidence for protein translocation along DNA
Article Snippet: Plasmid pDERI1 was generated by deletion of the Ssp I– Sap I fragment from plasmid pUC19, resulting in a plasmid 1,821 bp in length. .. Singly nicked plasmid DNA was produced in a 30-μl reaction mixture containing 0.5 μg of DNA, 20 mM Tris⋅HCl (pH 7.5), 50 mM NaCl, 10 mM MgCl2 , 360 μg/ml ethidium bromide, and 1 μg/ml DNase I at 30°C for 30 min. .. The reaction was stopped by the addition of 0.1 vol of 5% (wt/vol) SDS/50 mM EDTA/30 μg/ml proteinase K and subsequent incubation at 65°C for 30 min. DNA was purified by extraction with phenol and phenol/chloroform (1:1, vol/vol), precipitated with ethanol, and dissolved in 10 mM Tris⋅HCl (pH 8.0)/1 mM EDTA.

Mouse Assay:

Article Title: DNA Methyltransferase 3a and Mitogen-activated Protein Kinase Signaling Regulate the Expression of Fibroblast Growth Factor-inducible 14 (Fn14) during Denervation-induced Skeletal Muscle Atrophy *
Article Snippet: In brief, plasmids were prepared using an endotoxin-free kit (Qiagen) and suspended in sterile saline solution. .. The mice were anesthetized, and a small portion of TA muscle of both hind limbs was exposed and injected with plasmid DNA (20 μg in 20 μl of saline). .. After 1 min of plasmid DNA injection, a pair of platinum plate electrodes was placed against the closely shaved skin on either side of the small surgical incision, and electric pulses were delivered transcutaneously.

Injection:

Article Title: DNA Methyltransferase 3a and Mitogen-activated Protein Kinase Signaling Regulate the Expression of Fibroblast Growth Factor-inducible 14 (Fn14) during Denervation-induced Skeletal Muscle Atrophy *
Article Snippet: In brief, plasmids were prepared using an endotoxin-free kit (Qiagen) and suspended in sterile saline solution. .. The mice were anesthetized, and a small portion of TA muscle of both hind limbs was exposed and injected with plasmid DNA (20 μg in 20 μl of saline). .. After 1 min of plasmid DNA injection, a pair of platinum plate electrodes was placed against the closely shaved skin on either side of the small surgical incision, and electric pulses were delivered transcutaneously.

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    ATUM pbr322 plasmid dna
    Electrophoresis mobility shift assays for cisplatin and heterometallic Ti–Au compounds 5a – d (see the Experimental Section for details). <t>DNA</t> refers to untreated plasmid <t>pBR322.</t> a , b , c , and d correspond to metal/DNA bp ratios of 0.25, 0.5, 1.0, and 2.0, respectively.
    Pbr322 Plasmid Dna, supplied by ATUM, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pbr322 plasmid dna/product/ATUM
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pbr322 plasmid dna - by Bioz Stars, 2021-03
    86/100 stars
      Buy from Supplier

    86
    ATUM hindiii linearized pryg plasmid dna
    Etoplatin-N2β, but not the N2α epimer, more potently inhibits the supercoil relaxation activity of Top2 by inducing the formation of ethylenediaminetetraacetic acid (EDTA)-resistant <t>DNA</t> breaks. ( A ) Inhibition of the relaxation activity of hTop2 by etoposide and etoplatins. Each relaxation reaction contains 300 ng of supercoiled (SC) <t>pRYG</t> plasmid DNA. The enzyme-positive reactions contain 80 ng of hTop2β △CTD (designated as hTop2β). OC stands for open circle (full-relaxed product produced by Top2) DNA. ( B ) The DNA cleavage assay shows the production of EDTA-resistant, hTop2-mediated DNA breaks in the presence of etoplatin-N2β. Each cleavage reaction contains 250 ng of <t>HindIII-linearized</t> (L) pRYG plasmid DNA. A total of 1.2 μg of hTop2β △CTD (upper panel, designated as hTop2β) or hTop2α △CTD (lower panel, designated as hTop2α) was added to the respective enzyme-positive reactions. To stop the cleavage reaction, Sodium dodecyl sulphate (SDS) and EDTA were added in the indicated order and the denatured enzyme was removed by proteinase K digestion. Lanes labeled with -ProK indicates no proteinase K treatment after the reaction was stopped. The disappearance and reemergence of the linear substrate DNA (L) indicate the production and resealing of DNA breaks, respectively. Refer to Supplementary Figure S2 for the full-sized image of this gel.
    Hindiii Linearized Pryg Plasmid Dna, supplied by ATUM, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hindiii linearized pryg plasmid dna/product/ATUM
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hindiii linearized pryg plasmid dna - by Bioz Stars, 2021-03
    86/100 stars
      Buy from Supplier

    86
    ATUM plasmid dna
    Southern analysis of <t>transposon</t> insertions into S. cerevisiae genome. Genomic <t>DNA</t> of 17 G418-resistant yeast clones (diploid strain FY1679) was digested with BamHI + BglII (left) or HindIII (right) and probed with labeled kanMX4 DNA. (Lanes 1–17) Insertion mutants, (lane C) Genomic DNA of the original FY1679 recipient strain as a negative control, (lane P) Plasmid DNA containing the kanMX4-Mu transposon digested with HindIII as a positive control, fragment sizes on the left.
    Plasmid Dna, supplied by ATUM, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmid dna/product/ATUM
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    plasmid dna - by Bioz Stars, 2021-03
    86/100 stars
      Buy from Supplier

    Image Search Results


    Electrophoresis mobility shift assays for cisplatin and heterometallic Ti–Au compounds 5a – d (see the Experimental Section for details). DNA refers to untreated plasmid pBR322. a , b , c , and d correspond to metal/DNA bp ratios of 0.25, 0.5, 1.0, and 2.0, respectively.

    Journal: Organometallics

    Article Title: Titanocene–Gold Complexes Containing N-Heterocyclic Carbene Ligands Inhibit Growth of Prostate, Renal, and Colon Cancers in Vitro

    doi: 10.1021/acs.organomet.6b00051

    Figure Lengend Snippet: Electrophoresis mobility shift assays for cisplatin and heterometallic Ti–Au compounds 5a – d (see the Experimental Section for details). DNA refers to untreated plasmid pBR322. a , b , c , and d correspond to metal/DNA bp ratios of 0.25, 0.5, 1.0, and 2.0, respectively.

    Article Snippet: Interactions of the New Compounds with Plasmid pBR322 (Gel Electrophoresis Mobility Shift Assay) Ten microliter aliquots of pBR322 plasmid DNA (20 μg/mL) in buffer (5 mM Tris/HCl, 50 mM NaClO4 , pH 7.39) were incubated with different concentrations of the compounds ( 4a – d , 5a – d , and cisplatin as control) (in the range 0.25 and 4.0 metal complex to DNA bp) at 37 °C for 20 h in the dark.

    Techniques: Electrophoresis, Mobility Shift, Plasmid Preparation

    Etoplatin-N2β, but not the N2α epimer, more potently inhibits the supercoil relaxation activity of Top2 by inducing the formation of ethylenediaminetetraacetic acid (EDTA)-resistant DNA breaks. ( A ) Inhibition of the relaxation activity of hTop2 by etoposide and etoplatins. Each relaxation reaction contains 300 ng of supercoiled (SC) pRYG plasmid DNA. The enzyme-positive reactions contain 80 ng of hTop2β △CTD (designated as hTop2β). OC stands for open circle (full-relaxed product produced by Top2) DNA. ( B ) The DNA cleavage assay shows the production of EDTA-resistant, hTop2-mediated DNA breaks in the presence of etoplatin-N2β. Each cleavage reaction contains 250 ng of HindIII-linearized (L) pRYG plasmid DNA. A total of 1.2 μg of hTop2β △CTD (upper panel, designated as hTop2β) or hTop2α △CTD (lower panel, designated as hTop2α) was added to the respective enzyme-positive reactions. To stop the cleavage reaction, Sodium dodecyl sulphate (SDS) and EDTA were added in the indicated order and the denatured enzyme was removed by proteinase K digestion. Lanes labeled with -ProK indicates no proteinase K treatment after the reaction was stopped. The disappearance and reemergence of the linear substrate DNA (L) indicate the production and resealing of DNA breaks, respectively. Refer to Supplementary Figure S2 for the full-sized image of this gel.

    Journal: Nucleic Acids Research

    Article Title: Producing irreversible topoisomerase II-mediated DNA breaks by site-specific Pt(II)-methionine coordination chemistry

    doi: 10.1093/nar/gkx742

    Figure Lengend Snippet: Etoplatin-N2β, but not the N2α epimer, more potently inhibits the supercoil relaxation activity of Top2 by inducing the formation of ethylenediaminetetraacetic acid (EDTA)-resistant DNA breaks. ( A ) Inhibition of the relaxation activity of hTop2 by etoposide and etoplatins. Each relaxation reaction contains 300 ng of supercoiled (SC) pRYG plasmid DNA. The enzyme-positive reactions contain 80 ng of hTop2β △CTD (designated as hTop2β). OC stands for open circle (full-relaxed product produced by Top2) DNA. ( B ) The DNA cleavage assay shows the production of EDTA-resistant, hTop2-mediated DNA breaks in the presence of etoplatin-N2β. Each cleavage reaction contains 250 ng of HindIII-linearized (L) pRYG plasmid DNA. A total of 1.2 μg of hTop2β △CTD (upper panel, designated as hTop2β) or hTop2α △CTD (lower panel, designated as hTop2α) was added to the respective enzyme-positive reactions. To stop the cleavage reaction, Sodium dodecyl sulphate (SDS) and EDTA were added in the indicated order and the denatured enzyme was removed by proteinase K digestion. Lanes labeled with -ProK indicates no proteinase K treatment after the reaction was stopped. The disappearance and reemergence of the linear substrate DNA (L) indicate the production and resealing of DNA breaks, respectively. Refer to Supplementary Figure S2 for the full-sized image of this gel.

    Article Snippet: The reaction was terminated by the addition of 0.4 μl of 0.5% sodium dodecyl sulphate (SDS) and 0.8 μl of 250 mM EDTA and incubated at 37 °C for 1 h. The DNA cleavage reaction (20 μl) was conducted at 37 °C for 30 min in a buffer composed of 1 mM ATP, 100 μg/ml BSA, 250 ng of HindIII-linearized pRYG plasmid DNA, 20 nM of indicated drug and 1.2 μg/μl of hTop2△CTD .

    Techniques: Activity Assay, Inhibition, Plasmid Preparation, Produced, DNA Cleavage Assay, Labeling

    Southern analysis of transposon insertions into S. cerevisiae genome. Genomic DNA of 17 G418-resistant yeast clones (diploid strain FY1679) was digested with BamHI + BglII (left) or HindIII (right) and probed with labeled kanMX4 DNA. (Lanes 1–17) Insertion mutants, (lane C) Genomic DNA of the original FY1679 recipient strain as a negative control, (lane P) Plasmid DNA containing the kanMX4-Mu transposon digested with HindIII as a positive control, fragment sizes on the left.

    Journal: Nucleic Acids Research

    Article Title: Bacteriophage Mu integration in yeast and mammalian genomes

    doi: 10.1093/nar/gkn801

    Figure Lengend Snippet: Southern analysis of transposon insertions into S. cerevisiae genome. Genomic DNA of 17 G418-resistant yeast clones (diploid strain FY1679) was digested with BamHI + BglII (left) or HindIII (right) and probed with labeled kanMX4 DNA. (Lanes 1–17) Insertion mutants, (lane C) Genomic DNA of the original FY1679 recipient strain as a negative control, (lane P) Plasmid DNA containing the kanMX4-Mu transposon digested with HindIII as a positive control, fragment sizes on the left.

    Article Snippet: An aliquot (1–2 μl) of transpososome preparation (containing 1 μg transposon DNA) or plasmid DNA (20 ng) was added to the cell suspension.

    Techniques: Clone Assay, Labeling, Negative Control, Plasmid Preparation, Positive Control

    Analysis of transpososome-mediated gene delivery into human ES cells. ( A ) Expression of eGFP. Human ES cell line FES29 was electroporated with Puro-eGFP-Mu transposons and selected for 2 days with puromycin. Surviving fluorescent colonies were isolated and further cultured as clonal cell lines for several passages. Most of the colonies of the clonal isolates showed uniform GFP expression. Two example clones are shown in the phase contrast (left) and fluorescent (right) micrographs. ( B ) Southern analysis of the insertions into the hES cell genome. Genomic DNA of nine puromycin-resistant hES cell clones was digested with BglII (left) or EcoRI (right) and probed with labeled Puro-eGFP-Mu transposon DNA. (Lane P) Undigested genomic DNA of clone 2 spiked with transposon DNA as a positive control. (M) Size marker.

    Journal: Nucleic Acids Research

    Article Title: Bacteriophage Mu integration in yeast and mammalian genomes

    doi: 10.1093/nar/gkn801

    Figure Lengend Snippet: Analysis of transpososome-mediated gene delivery into human ES cells. ( A ) Expression of eGFP. Human ES cell line FES29 was electroporated with Puro-eGFP-Mu transposons and selected for 2 days with puromycin. Surviving fluorescent colonies were isolated and further cultured as clonal cell lines for several passages. Most of the colonies of the clonal isolates showed uniform GFP expression. Two example clones are shown in the phase contrast (left) and fluorescent (right) micrographs. ( B ) Southern analysis of the insertions into the hES cell genome. Genomic DNA of nine puromycin-resistant hES cell clones was digested with BglII (left) or EcoRI (right) and probed with labeled Puro-eGFP-Mu transposon DNA. (Lane P) Undigested genomic DNA of clone 2 spiked with transposon DNA as a positive control. (M) Size marker.

    Article Snippet: An aliquot (1–2 μl) of transpososome preparation (containing 1 μg transposon DNA) or plasmid DNA (20 ng) was added to the cell suspension.

    Techniques: Expressing, Isolation, Cell Culture, Clone Assay, Labeling, Positive Control, Marker

    Southern blot analysis of transposon insertions into HeLa cell genome. Chromosomal DNA was doubly digested with BamHI + BglII and probed with labeled Kan/Neo-p15A-Mu transposon DNA. A total of 19 different G418-resistant clones are analyzed, some with their siblings (bracketed) for the verification of clonality. (Lane C) Genomic DNA of the recipient HeLa cell line as a negative control. (Lane P) HeLa cell DNA spiked with transposon DNA as a positive control. (Lane M) Size marker.

    Journal: Nucleic Acids Research

    Article Title: Bacteriophage Mu integration in yeast and mammalian genomes

    doi: 10.1093/nar/gkn801

    Figure Lengend Snippet: Southern blot analysis of transposon insertions into HeLa cell genome. Chromosomal DNA was doubly digested with BamHI + BglII and probed with labeled Kan/Neo-p15A-Mu transposon DNA. A total of 19 different G418-resistant clones are analyzed, some with their siblings (bracketed) for the verification of clonality. (Lane C) Genomic DNA of the recipient HeLa cell line as a negative control. (Lane P) HeLa cell DNA spiked with transposon DNA as a positive control. (Lane M) Size marker.

    Article Snippet: An aliquot (1–2 μl) of transpososome preparation (containing 1 μg transposon DNA) or plasmid DNA (20 ng) was added to the cell suspension.

    Techniques: Southern Blot, Labeling, Clone Assay, Negative Control, Positive Control, Marker

    Analysis of transpososome-mediated gene delivery into mouse ES cells. ( A ) Efficiency. Pre-assembled transpososomes made with Kan/Neo-p15A-Mu transposon were introduced into mouse cells by electroporation. Following G418 selection, surviving cell colonies were stained with methylene blue. Gene delivery was analyzed using transpososomes made with wild-type MuA protein (left) and active site mutant MuA(E392Q) (middle). Analysis was done also with linear transposon DNA (right). ( B ) Southern analysis of transposon insertions into the mES cell genome. Genomic DNA of 17 G418-resistant mES cell clones was doubly digested with BamHI + BglII and probed with labeled Kan/Neo-p15A-Mu transposon DNA. (Lanes 1–17) Transposon insertion mutants. (Lane C) Genomic DNA of the original AB2.2 recipient strain as a negative control. (Lane P) AB2.2 genomic DNA spiked with transposon DNA as a positive control. (Lane M) Size marker. The cross-hybridizing band present in all genomic DNA samples served as a loading control.

    Journal: Nucleic Acids Research

    Article Title: Bacteriophage Mu integration in yeast and mammalian genomes

    doi: 10.1093/nar/gkn801

    Figure Lengend Snippet: Analysis of transpososome-mediated gene delivery into mouse ES cells. ( A ) Efficiency. Pre-assembled transpososomes made with Kan/Neo-p15A-Mu transposon were introduced into mouse cells by electroporation. Following G418 selection, surviving cell colonies were stained with methylene blue. Gene delivery was analyzed using transpososomes made with wild-type MuA protein (left) and active site mutant MuA(E392Q) (middle). Analysis was done also with linear transposon DNA (right). ( B ) Southern analysis of transposon insertions into the mES cell genome. Genomic DNA of 17 G418-resistant mES cell clones was doubly digested with BamHI + BglII and probed with labeled Kan/Neo-p15A-Mu transposon DNA. (Lanes 1–17) Transposon insertion mutants. (Lane C) Genomic DNA of the original AB2.2 recipient strain as a negative control. (Lane P) AB2.2 genomic DNA spiked with transposon DNA as a positive control. (Lane M) Size marker. The cross-hybridizing band present in all genomic DNA samples served as a loading control.

    Article Snippet: An aliquot (1–2 μl) of transpososome preparation (containing 1 μg transposon DNA) or plasmid DNA (20 ng) was added to the cell suspension.

    Techniques: Electroporation, Selection, Staining, Mutagenesis, Clone Assay, Labeling, Negative Control, Positive Control, Marker

    DNA treatment impairs KLH antigen presentation. Peritoneal macrophages were incubated with DNA (20 µg) 72 h, 48 h, 24 h or 0 h prior to the treatment with KLH (100 µg). After KLH treatment (24 h), the peritoneal macrophages were fixed and CD4 T cells specific for KLH were added to the culture. Proliferation was measured after 72 h. Concanavalin A (40 µg/ml) was used as positive control. * p

    Journal: PLoS ONE

    Article Title: Endocytosis of DNA-Hsp65 Alters the pH of the Late Endosome/Lysosome and Interferes with Antigen Presentation

    doi: 10.1371/journal.pone.0000923

    Figure Lengend Snippet: DNA treatment impairs KLH antigen presentation. Peritoneal macrophages were incubated with DNA (20 µg) 72 h, 48 h, 24 h or 0 h prior to the treatment with KLH (100 µg). After KLH treatment (24 h), the peritoneal macrophages were fixed and CD4 T cells specific for KLH were added to the culture. Proliferation was measured after 72 h. Concanavalin A (40 µg/ml) was used as positive control. * p

    Article Snippet: The results showed that plasmid DNA completely inhibited the KLH antigen presentation when the DNA (20 µg) was added to the culture simultaneously with KLH, and antigen presentation was impaired when DNA was added at 24 h, 48 h or 72 h before exposure to KLH.

    Techniques: Incubation, Positive Control