standard miniprep kit  (Qiagen)


Bioz Verified Symbol Qiagen is a verified supplier
Bioz Manufacturer Symbol Qiagen manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90

    Structured Review

    Qiagen standard miniprep kit
    Confirmation of HGT from replicating and homology plasmids. ( a ) plasmid DNA isolated with a standard <t>miniprep</t> kit and digested using SspI. Lanes 1–3: Kanamycin-resistant Acinetobacter clones resulting from HGT of pBAV1k. Lanes 4–6: Kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03+homology. Lane 7: E. coli carrying pBAV1k. ( b ) inverse PCR with primers located in the kan gene using genomic DNA isolated from kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03H.
    Standard Miniprep Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/standard miniprep kit/product/Qiagen
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    standard miniprep kit - by Bioz Stars, 2020-04
    90/100 stars

    Images

    1) Product Images from "Inter-species population dynamics enhance microbial horizontal gene transfer and spread of antibiotic resistance"

    Article Title: Inter-species population dynamics enhance microbial horizontal gene transfer and spread of antibiotic resistance

    Journal: eLife

    doi: 10.7554/eLife.25950

    Confirmation of HGT from replicating and homology plasmids. ( a ) plasmid DNA isolated with a standard miniprep kit and digested using SspI. Lanes 1–3: Kanamycin-resistant Acinetobacter clones resulting from HGT of pBAV1k. Lanes 4–6: Kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03+homology. Lane 7: E. coli carrying pBAV1k. ( b ) inverse PCR with primers located in the kan gene using genomic DNA isolated from kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03H.
    Figure Legend Snippet: Confirmation of HGT from replicating and homology plasmids. ( a ) plasmid DNA isolated with a standard miniprep kit and digested using SspI. Lanes 1–3: Kanamycin-resistant Acinetobacter clones resulting from HGT of pBAV1k. Lanes 4–6: Kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03+homology. Lane 7: E. coli carrying pBAV1k. ( b ) inverse PCR with primers located in the kan gene using genomic DNA isolated from kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03H.

    Techniques Used: Plasmid Preparation, Isolation, Clone Assay, Inverse PCR

    2) Product Images from "Inter-species population dynamics enhance microbial horizontal gene transfer and spread of antibiotic resistance"

    Article Title: Inter-species population dynamics enhance microbial horizontal gene transfer and spread of antibiotic resistance

    Journal: eLife

    doi: 10.7554/eLife.25950

    Confirmation of HGT from replicating and homology plasmids. ( a ) plasmid DNA isolated with a standard miniprep kit and digested using SspI. Lanes 1–3: Kanamycin-resistant Acinetobacter clones resulting from HGT of pBAV1k. Lanes 4–6: Kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03+homology. Lane 7: E. coli carrying pBAV1k. ( b ) inverse PCR with primers located in the kan gene using genomic DNA isolated from kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03H.
    Figure Legend Snippet: Confirmation of HGT from replicating and homology plasmids. ( a ) plasmid DNA isolated with a standard miniprep kit and digested using SspI. Lanes 1–3: Kanamycin-resistant Acinetobacter clones resulting from HGT of pBAV1k. Lanes 4–6: Kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03+homology. Lane 7: E. coli carrying pBAV1k. ( b ) inverse PCR with primers located in the kan gene using genomic DNA isolated from kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03H.

    Techniques Used: Plasmid Preparation, Isolation, Clone Assay, Inverse PCR

    Related Articles

    Clone Assay:

    Article Title: Modifiers of C9orf72 DPR toxicity implicate nucleocytoplasmic transport impairments in c9FTD/ALS
    Article Snippet: .. Plasmid DNA from the expression clones were isolated using the Plasmid Plus 96 miniprep kit (Qiagen). .. DNA was dried in individual wells of 96-well microtiter plates and transformed into a strain expressing (PR)50 integrated at the HIS3 locus.

    Article Title: Biology and interactions of two distinct monopartite begomoviruses and betasatellites associated with radish leaf curl disease in India
    Article Snippet: .. Sequence comparisons Putative full-length clones were purified using a Qiagen plasmid miniprep kit (Qiagen) and their sequences were determined commercially (Chromous Biotech, India). .. The sequence results were analyzed by comparing the sequence to other begomovirus sequences in GenBank using BLAST, followed by analysis using MEGA program version 4 ( http://www.megasoftware.net/mega4/features.html ) [ ] with default parameters.

    Article Title: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite
    Article Snippet: To improve genome coverage, the arrayed P. berghei genomic DNA library in the pJAZZ-OK linear plasmid (Lucigen) that provides the starting point for vector production was doubled in size to ∼10,000 clones. .. Intermediate vector DNA was obtained using a QIAGEN Plasmid Plus 96 Miniprep kit and eluted into 65 μl Tris-EDTA buffer.

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution. .. Plasmids from at least two different clones of each fragment, for each specimen, were bi-directionally sequenced on a LiCor 4200 L long-read, dual-laser sequencer.

    Article Title: An atlas of active enhancers across human cell types and tissues
    Article Snippet: We inserted an EF1α basal promoter fragment into Hin dIII and Nhe I sites of the multiple cloning site in pGL4.10 (Promega) to construct a basal pGL4.10EF1α backbone. .. Vectors were purified using the QIAGEN Plasmid Plus 96 Miniprep Kit (QIAGEN).

    Article Title: Capped antigenomic RNA transcript facilitates rescue of a plant rhabdovirus
    Article Snippet: .. General Plasmid DNA was extracted using an AxyPrep plasmid miniprep kit (Axygen, Union, CA) or a QIAGEN plasmid miniprep kit (QIAGEN, Hilden, Germany) and used for cloning and sequencing. .. DNAs for subcloning were recovered from gel slices using an AxyPrep DNA gel extraction kit (Axygen, Union, CA) or a Purelink Quick gel extraction kit (Invitrogen, Shanghai, China).

    Article Title: Functional and phylogenetic characterization of noncanonical vitamin B12–binding proteins in zebrafish suggests involvement in cobalamin transport
    Article Snippet: .. Potential destination clones were isolated according to the protocol of the Qiagen Plasmid Miniprep kit (Qiagen) for further use. .. Chinese Hamster Ovarian K1 cells were obtained from liquid nitrogen stocks, thawed, and plated in liquid media consisting of DMEM supplemented with 10% FBS (Thermo Fisher, lot no.: 1901903), 1× Non-Essential Amino Acids (Thermo Fisher), and 2 m m l -glutamine (Thermo Fisher).

    Amplification:

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: Forward and reverse primers were used for PCR amplification ( ). .. The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit.

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: Paragraph title: DNA isolation amplification and sequencing ... Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution.

    Article Title: Novel Biocatalysts Based on S-Layer Self-Assembly of Geobacillus Stearothermophilus NRS 2004/3a: A Nanobiotechnological Approach
    Article Snippet: Plasmid DNA from transformed cells was isolated with the Qiagen Plasmid miniprep kit. .. PCR conditions were optimized for each primer pair and amplification products were purified using the MinElute PCR purification kit.

    Positive Control:

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: CRITICAL STEP Wells G12 and H12 are used as negative control (water as template) and positive control (10 ng of empty pDEST-AD), respectively. .. 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions.

    Construct:

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: The pGEM7Zf(+)- fbc B plasmid ( ) was constructed by ligating the NsiI-XbaI fragment from pRKD418- fbc FBC6H Q into the NsiI and XbaI sites of the pGEM7Zf(+) plasmid. .. The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit.

    Article Title: An atlas of active enhancers across human cell types and tissues
    Article Snippet: We inserted an EF1α basal promoter fragment into Hin dIII and Nhe I sites of the multiple cloning site in pGL4.10 (Promega) to construct a basal pGL4.10EF1α backbone. .. Vectors were purified using the QIAGEN Plasmid Plus 96 Miniprep Kit (QIAGEN).

    Incubation:

    Article Title: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite
    Article Snippet: After two rounds of antibiotic selection, cultures were reinoculated into fresh selective medium and incubated for no longer than 16 hr at 37°C to favor the correct recombination product. .. Intermediate vector DNA was obtained using a QIAGEN Plasmid Plus 96 Miniprep kit and eluted into 65 μl Tris-EDTA buffer.

    Expressing:

    Article Title: Modifiers of C9orf72 DPR toxicity implicate nucleocytoplasmic transport impairments in c9FTD/ALS
    Article Snippet: .. Plasmid DNA from the expression clones were isolated using the Plasmid Plus 96 miniprep kit (Qiagen). .. DNA was dried in individual wells of 96-well microtiter plates and transformed into a strain expressing (PR)50 integrated at the HIS3 locus.

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: .. The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit. .. Because R. sphaeroides cells contain four types of endogenous plasmids, the isolated plasmids lacked the purity and concentration needed for direct sequencing.

    Genome Wide:

    Article Title: Modifiers of C9orf72 DPR toxicity implicate nucleocytoplasmic transport impairments in c9FTD/ALS
    Article Snippet: Yeast Genetic screens The yeast FLEXGene collection was used to perform the genome-wide plasmid overexpression screen . .. Plasmid DNA from the expression clones were isolated using the Plasmid Plus 96 miniprep kit (Qiagen).

    Modification:

    Article Title: Modifiers of C9orf72 DPR toxicity implicate nucleocytoplasmic transport impairments in c9FTD/ALS
    Article Snippet: Plasmid DNA from the expression clones were isolated using the Plasmid Plus 96 miniprep kit (Qiagen). .. A standard lithium acetate transformation protocol was modified for automation and used by employing a BIOROBOT Rapidplate 96-well pipettor (Qiagen).

    Article Title: An atlas of active enhancers across human cell types and tissues
    Article Snippet: Five μl of purified PCR products were ligated with 100 ng of the BamH I- and Sal I-digested modified pGL4.10EF1α and pGL4.10 plasmids using Ligation-high (TOYOBO), and transformed into DH5α competent cells (TOYOBO). .. Vectors were purified using the QIAGEN Plasmid Plus 96 Miniprep Kit (QIAGEN).

    Western Blot:

    Article Title: Novel Biocatalysts Based on S-Layer Self-Assembly of Geobacillus Stearothermophilus NRS 2004/3a: A Nanobiotechnological Approach
    Article Snippet: Western immunoblotting using anti-His-tag monoclonal antibody (Novagen, Madison, WI), anti-SgsE antibody and anti-RmlA antibody, was done as described elsewhere. .. Plasmid DNA from transformed cells was isolated with the Qiagen Plasmid miniprep kit.

    Transformation Assay:

    Article Title: Identification of an Archaeal Type II Isopentenyl Diphosphate Isomerase in Methanothermobacter thermautotrophicus
    Article Snippet: Minipreparations of plasmid DNA for restriction analysis were obtained by using a Qiagen plasmid miniprep kit. .. Restriction digestion, ligation, and transformation of competent E. coli and S. enterica serovar Typhimurium cells were conducted as described by Sambrook et al. ( ) PCR was performed using the polymerase mix provided in the Advantage PCR kit from Clontech.

    Article Title: Modifiers of C9orf72 DPR toxicity implicate nucleocytoplasmic transport impairments in c9FTD/ALS
    Article Snippet: Plasmid DNA from the expression clones were isolated using the Plasmid Plus 96 miniprep kit (Qiagen). .. DNA was dried in individual wells of 96-well microtiter plates and transformed into a strain expressing (PR)50 integrated at the HIS3 locus.

    Article Title: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite
    Article Snippet: Intermediate vector DNA was obtained using a QIAGEN Plasmid Plus 96 Miniprep kit and eluted into 65 μl Tris-EDTA buffer. .. Electrocompetent E. coli TSA were transformed and plated on YEG-Cl agar containing kanamycin, and four colonies were picked to verify the sequence of their homology arms.

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: Transformed cells were plated on kanamycin/X-gal plates for blue/white selection. .. Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution.

    Article Title: An atlas of active enhancers across human cell types and tissues
    Article Snippet: Five μl of purified PCR products were ligated with 100 ng of the BamH I- and Sal I-digested modified pGL4.10EF1α and pGL4.10 plasmids using Ligation-high (TOYOBO), and transformed into DH5α competent cells (TOYOBO). .. Vectors were purified using the QIAGEN Plasmid Plus 96 Miniprep Kit (QIAGEN).

    Article Title: Novel Biocatalysts Based on S-Layer Self-Assembly of Geobacillus Stearothermophilus NRS 2004/3a: A Nanobiotechnological Approach
    Article Snippet: .. Plasmid DNA from transformed cells was isolated with the Qiagen Plasmid miniprep kit. ..

    Over Expression:

    Article Title: Modifiers of C9orf72 DPR toxicity implicate nucleocytoplasmic transport impairments in c9FTD/ALS
    Article Snippet: Yeast Genetic screens The yeast FLEXGene collection was used to perform the genome-wide plasmid overexpression screen . .. Plasmid DNA from the expression clones were isolated using the Plasmid Plus 96 miniprep kit (Qiagen).

    Gel Purification:

    Article Title: Identification of an Archaeal Type II Isopentenyl Diphosphate Isomerase in Methanothermobacter thermautotrophicus
    Article Snippet: Minipreparations of plasmid DNA for restriction analysis were obtained by using a Qiagen plasmid miniprep kit. .. DNA fragments were purified in agarose gels (Bio-Rad) using a gel purification kit from Qiagen.

    Ligation:

    Article Title: Identification of an Archaeal Type II Isopentenyl Diphosphate Isomerase in Methanothermobacter thermautotrophicus
    Article Snippet: Minipreparations of plasmid DNA for restriction analysis were obtained by using a Qiagen plasmid miniprep kit. .. Restriction digestion, ligation, and transformation of competent E. coli and S. enterica serovar Typhimurium cells were conducted as described by Sambrook et al. ( ) PCR was performed using the polymerase mix provided in the Advantage PCR kit from Clontech.

    Article Title: An atlas of active enhancers across human cell types and tissues
    Article Snippet: Five μl of purified PCR products were ligated with 100 ng of the BamH I- and Sal I-digested modified pGL4.10EF1α and pGL4.10 plasmids using Ligation-high (TOYOBO), and transformed into DH5α competent cells (TOYOBO). .. Vectors were purified using the QIAGEN Plasmid Plus 96 Miniprep Kit (QIAGEN).

    Cell Culture:

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: White colonies were picked and cultured overnight in Luria-Bertani broth. .. Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution.

    Article Title: An atlas of active enhancers across human cell types and tissues
    Article Snippet: Vectors were purified using the QIAGEN Plasmid Plus 96 Miniprep Kit (QIAGEN). .. HeLa-S3 cells (JCRB Cell Bank) were cultured in MEM (WAKO) supplemented with 10% FBS (NICHIREI BIOSCIENCE INC., Lot No. 7G0031), 100 Units/mL penicillin and 100 μg/mL streptomycin (both Life Technologies).

    Generated:

    Article Title: Biology and interactions of two distinct monopartite begomoviruses and betasatellites associated with radish leaf curl disease in India
    Article Snippet: Sequence comparisons Putative full-length clones were purified using a Qiagen plasmid miniprep kit (Qiagen) and their sequences were determined commercially (Chromous Biotech, India). .. The full-length genome sequences of representative begomoviruses and betasatellites were aligned using ClustalW and dendogram was generated using 1000 bootstrap repetitions.

    DNA Sequencing:

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: The presence of the engineered mutations was confirmed by DNA sequencing before and after semi-aerobic growth of the cells. .. The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit.

    Protein Concentration:

    Article Title: Novel Biocatalysts Based on S-Layer Self-Assembly of Geobacillus Stearothermophilus NRS 2004/3a: A Nanobiotechnological Approach
    Article Snippet: [ ] Protein concentration was determined by the Bio-Rad protein assay (Bio-Rad, Vienna, Austria) using bovine serum albumin (BSA) as standard. .. Plasmid DNA from transformed cells was isolated with the Qiagen Plasmid miniprep kit.

    Sequencing:

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit. .. Because R. sphaeroides cells contain four types of endogenous plasmids, the isolated plasmids lacked the purity and concentration needed for direct sequencing.

    Article Title: Biology and interactions of two distinct monopartite begomoviruses and betasatellites associated with radish leaf curl disease in India
    Article Snippet: .. Sequence comparisons Putative full-length clones were purified using a Qiagen plasmid miniprep kit (Qiagen) and their sequences were determined commercially (Chromous Biotech, India). .. The sequence results were analyzed by comparing the sequence to other begomovirus sequences in GenBank using BLAST, followed by analysis using MEGA program version 4 ( http://www.megasoftware.net/mega4/features.html ) [ ] with default parameters.

    Article Title: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite
    Article Snippet: Intermediate vector DNA was obtained using a QIAGEN Plasmid Plus 96 Miniprep kit and eluted into 65 μl Tris-EDTA buffer. .. Electrocompetent E. coli TSA were transformed and plated on YEG-Cl agar containing kanamycin, and four colonies were picked to verify the sequence of their homology arms.

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: Paragraph title: DNA isolation amplification and sequencing ... Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution.

    Article Title: Identification of fasciclin-like arabinogalactan proteins in textile hemp (Cannabis sativa L.): in silico analyses and gene expression patterns in different tissues
    Article Snippet: .. Plasmids were extracted using the QIAGEN plasmid miniprep kit and sequenced on an Applied Biosystems 3500 Genetic Analyser using the BigDye Terminator v3.1 Cycle Sequencing and the BigDye XTerminator Purification kits, according to the manufacturer’s instructions. ..

    Article Title: Capped antigenomic RNA transcript facilitates rescue of a plant rhabdovirus
    Article Snippet: .. General Plasmid DNA was extracted using an AxyPrep plasmid miniprep kit (Axygen, Union, CA) or a QIAGEN plasmid miniprep kit (QIAGEN, Hilden, Germany) and used for cloning and sequencing. .. DNAs for subcloning were recovered from gel slices using an AxyPrep DNA gel extraction kit (Axygen, Union, CA) or a Purelink Quick gel extraction kit (Invitrogen, Shanghai, China).

    Article Title: Functional and phylogenetic characterization of noncanonical vitamin B12–binding proteins in zebrafish suggests involvement in cobalamin transport
    Article Snippet: Entry clone sequencing demonstrated that all plasmid sequences were as expected. .. Potential destination clones were isolated according to the protocol of the Qiagen Plasmid Miniprep kit (Qiagen) for further use.

    Molecular Weight:

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: Bands of the expected molecular weight were excised, purified and ligated into a plasmid vector using Topo-TA kits (Invitrogen). .. Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution.

    DNA Extraction:

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: Paragraph title: DNA isolation amplification and sequencing ... Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution.

    Marker:

    Article Title: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite
    Article Snippet: The PCR product consisting of a zeo-pheS marker and 50 bp primer extensions homologous to the gene of interest was purified using the High Pure 96 UF Cleanup Kit (Roche) to improve lambda Red recombination efficiency in E. coli . .. Intermediate vector DNA was obtained using a QIAGEN Plasmid Plus 96 Miniprep kit and eluted into 65 μl Tris-EDTA buffer.

    Radioactivity:

    Article Title: Identification of an Archaeal Type II Isopentenyl Diphosphate Isomerase in Methanothermobacter thermautotrophicus
    Article Snippet: Minipreparations of plasmid DNA for restriction analysis were obtained by using a Qiagen plasmid miniprep kit. .. Radioactivity was measured in Cytoscint scintillation fluid (ICN Biomedicals).

    Mutagenesis:

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit. .. Therefore, a 2-kilobase pair DNA segment containing the mutation sequence was amplified from the isolated plasmids by PCR.

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: Paragraph title: Preparation of mutagenesis PCR templates • TIMING 3 d ... 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions.

    Isolation:

    Article Title: Modifiers of C9orf72 DPR toxicity implicate nucleocytoplasmic transport impairments in c9FTD/ALS
    Article Snippet: .. Plasmid DNA from the expression clones were isolated using the Plasmid Plus 96 miniprep kit (Qiagen). .. DNA was dried in individual wells of 96-well microtiter plates and transformed into a strain expressing (PR)50 integrated at the HIS3 locus.

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit. .. Because R. sphaeroides cells contain four types of endogenous plasmids, the isolated plasmids lacked the purity and concentration needed for direct sequencing.

    Article Title: Novel Biocatalysts Based on S-Layer Self-Assembly of Geobacillus Stearothermophilus NRS 2004/3a: A Nanobiotechnological Approach
    Article Snippet: .. Plasmid DNA from transformed cells was isolated with the Qiagen Plasmid miniprep kit. ..

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: .. 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions. .. Measure DNA concentration by a high-throughput NanoDrop 8000 spectrophotometer.

    Article Title: Functional and phylogenetic characterization of noncanonical vitamin B12–binding proteins in zebrafish suggests involvement in cobalamin transport
    Article Snippet: .. Potential destination clones were isolated according to the protocol of the Qiagen Plasmid Miniprep kit (Qiagen) for further use. .. Chinese Hamster Ovarian K1 cells were obtained from liquid nitrogen stocks, thawed, and plated in liquid media consisting of DMEM supplemented with 10% FBS (Thermo Fisher, lot no.: 1901903), 1× Non-Essential Amino Acids (Thermo Fisher), and 2 m m l -glutamine (Thermo Fisher).

    Subcloning:

    Article Title: Capped antigenomic RNA transcript facilitates rescue of a plant rhabdovirus
    Article Snippet: General Plasmid DNA was extracted using an AxyPrep plasmid miniprep kit (Axygen, Union, CA) or a QIAGEN plasmid miniprep kit (QIAGEN, Hilden, Germany) and used for cloning and sequencing. .. DNAs for subcloning were recovered from gel slices using an AxyPrep DNA gel extraction kit (Axygen, Union, CA) or a Purelink Quick gel extraction kit (Invitrogen, Shanghai, China).

    Labeling:

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution. .. SequiTherm Excel ™ sequencing kits (Epicentre Technologies) and infrared labeled primers were used for sequencing reactions.

    Purification:

    Article Title: Identification of an Archaeal Type II Isopentenyl Diphosphate Isomerase in Methanothermobacter thermautotrophicus
    Article Snippet: Minipreparations of plasmid DNA for restriction analysis were obtained by using a Qiagen plasmid miniprep kit. .. DNA fragments were purified in agarose gels (Bio-Rad) using a gel purification kit from Qiagen.

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: .. The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit. .. Because R. sphaeroides cells contain four types of endogenous plasmids, the isolated plasmids lacked the purity and concentration needed for direct sequencing.

    Article Title: Biology and interactions of two distinct monopartite begomoviruses and betasatellites associated with radish leaf curl disease in India
    Article Snippet: .. Sequence comparisons Putative full-length clones were purified using a Qiagen plasmid miniprep kit (Qiagen) and their sequences were determined commercially (Chromous Biotech, India). .. The sequence results were analyzed by comparing the sequence to other begomovirus sequences in GenBank using BLAST, followed by analysis using MEGA program version 4 ( http://www.megasoftware.net/mega4/features.html ) [ ] with default parameters.

    Article Title: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite
    Article Snippet: The PCR product consisting of a zeo-pheS marker and 50 bp primer extensions homologous to the gene of interest was purified using the High Pure 96 UF Cleanup Kit (Roche) to improve lambda Red recombination efficiency in E. coli . .. Intermediate vector DNA was obtained using a QIAGEN Plasmid Plus 96 Miniprep kit and eluted into 65 μl Tris-EDTA buffer.

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: .. Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution. .. SequiTherm Excel ™ sequencing kits (Epicentre Technologies) and infrared labeled primers were used for sequencing reactions.

    Article Title: An atlas of active enhancers across human cell types and tissues
    Article Snippet: .. Vectors were purified using the QIAGEN Plasmid Plus 96 Miniprep Kit (QIAGEN). .. HeLa-S3 cells (JCRB Cell Bank) were cultured in MEM (WAKO) supplemented with 10% FBS (NICHIREI BIOSCIENCE INC., Lot No. 7G0031), 100 Units/mL penicillin and 100 μg/mL streptomycin (both Life Technologies).

    Article Title: Identification of fasciclin-like arabinogalactan proteins in textile hemp (Cannabis sativa L.): in silico analyses and gene expression patterns in different tissues
    Article Snippet: .. Plasmids were extracted using the QIAGEN plasmid miniprep kit and sequenced on an Applied Biosystems 3500 Genetic Analyser using the BigDye Terminator v3.1 Cycle Sequencing and the BigDye XTerminator Purification kits, according to the manufacturer’s instructions. ..

    Article Title: Novel Biocatalysts Based on S-Layer Self-Assembly of Geobacillus Stearothermophilus NRS 2004/3a: A Nanobiotechnological Approach
    Article Snippet: Plasmid DNA from transformed cells was isolated with the Qiagen Plasmid miniprep kit. .. PCR conditions were optimized for each primer pair and amplification products were purified using the MinElute PCR purification kit.

    Polymerase Chain Reaction:

    Article Title: Identification of an Archaeal Type II Isopentenyl Diphosphate Isomerase in Methanothermobacter thermautotrophicus
    Article Snippet: Minipreparations of plasmid DNA for restriction analysis were obtained by using a Qiagen plasmid miniprep kit. .. Restriction digestion, ligation, and transformation of competent E. coli and S. enterica serovar Typhimurium cells were conducted as described by Sambrook et al. ( ) PCR was performed using the polymerase mix provided in the Advantage PCR kit from Clontech.

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: Forward and reverse primers were used for PCR amplification ( ). .. The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit.

    Article Title: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite
    Article Snippet: The PCR product consisting of a zeo-pheS marker and 50 bp primer extensions homologous to the gene of interest was purified using the High Pure 96 UF Cleanup Kit (Roche) to improve lambda Red recombination efficiency in E. coli . .. Intermediate vector DNA was obtained using a QIAGEN Plasmid Plus 96 Miniprep kit and eluted into 65 μl Tris-EDTA buffer.

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: PCR products were electrophoresed on 1% agarose gels. .. Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution.

    Article Title: An atlas of active enhancers across human cell types and tissues
    Article Snippet: Correct insertion of the PCR products into the plasmids was checked by colony PCR. .. Vectors were purified using the QIAGEN Plasmid Plus 96 Miniprep Kit (QIAGEN).

    Article Title: Identification of fasciclin-like arabinogalactan proteins in textile hemp (Cannabis sativa L.): in silico analyses and gene expression patterns in different tissues
    Article Snippet: PCR products were ligated into the pGEM-T Easy vector, following the manufacturer’s instructions and cloned into JM109 chemically competent cells. .. Plasmids were extracted using the QIAGEN plasmid miniprep kit and sequenced on an Applied Biosystems 3500 Genetic Analyser using the BigDye Terminator v3.1 Cycle Sequencing and the BigDye XTerminator Purification kits, according to the manufacturer’s instructions.

    Article Title: Capped antigenomic RNA transcript facilitates rescue of a plant rhabdovirus
    Article Snippet: General Plasmid DNA was extracted using an AxyPrep plasmid miniprep kit (Axygen, Union, CA) or a QIAGEN plasmid miniprep kit (QIAGEN, Hilden, Germany) and used for cloning and sequencing. .. Oligonucleotides used for PCR amplifications were purchased from Invitrogen (Shanghai, China) or TsingKe (Hangzhou, China), and amplifications were performed using a high-fidelity KOD-Plus-Neo DNA polymerase (Toyobo, Osaka, Japan) or Phusion DNA polymerase (New England BioLabs, Shanghai, China).

    Article Title: Novel Biocatalysts Based on S-Layer Self-Assembly of Geobacillus Stearothermophilus NRS 2004/3a: A Nanobiotechnological Approach
    Article Snippet: Plasmid DNA from transformed cells was isolated with the Qiagen Plasmid miniprep kit. .. [ ] Polymerase chain reaction (PCR Sprint thermocycler, Hybaid, Ashford, UK) was performed using Pwo polymerase (Roche, Vienna, Austria).

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: Paragraph title: Preparation of mutagenesis PCR templates • TIMING 3 d ... 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions.

    Plasmid Preparation:

    Article Title: Identification of an Archaeal Type II Isopentenyl Diphosphate Isomerase in Methanothermobacter thermautotrophicus
    Article Snippet: .. Minipreparations of plasmid DNA for restriction analysis were obtained by using a Qiagen plasmid miniprep kit. .. DNA fragments were purified in agarose gels (Bio-Rad) using a gel purification kit from Qiagen.

    Article Title: Modifiers of C9orf72 DPR toxicity implicate nucleocytoplasmic transport impairments in c9FTD/ALS
    Article Snippet: .. Plasmid DNA from the expression clones were isolated using the Plasmid Plus 96 miniprep kit (Qiagen). .. DNA was dried in individual wells of 96-well microtiter plates and transformed into a strain expressing (PR)50 integrated at the HIS3 locus.

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: .. The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit. .. Because R. sphaeroides cells contain four types of endogenous plasmids, the isolated plasmids lacked the purity and concentration needed for direct sequencing.

    Article Title: Biology and interactions of two distinct monopartite begomoviruses and betasatellites associated with radish leaf curl disease in India
    Article Snippet: .. Sequence comparisons Putative full-length clones were purified using a Qiagen plasmid miniprep kit (Qiagen) and their sequences were determined commercially (Chromous Biotech, India). .. The sequence results were analyzed by comparing the sequence to other begomovirus sequences in GenBank using BLAST, followed by analysis using MEGA program version 4 ( http://www.megasoftware.net/mega4/features.html ) [ ] with default parameters.

    Article Title: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite
    Article Snippet: .. Intermediate vector DNA was obtained using a QIAGEN Plasmid Plus 96 Miniprep kit and eluted into 65 μl Tris-EDTA buffer. .. DNA purity proved critical for the Gateway reaction, which was set up in 20 μl using 2 μl LR clonase (Invitrogen), 100 ng Gateway entry plasmid (e.g., pR6K-attL1-3xHA-hdhfr-yfcu -attL2), LR clonase buffer, and 300 ng purified intermediate vector.

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: .. Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution. .. SequiTherm Excel ™ sequencing kits (Epicentre Technologies) and infrared labeled primers were used for sequencing reactions.

    Article Title: An atlas of active enhancers across human cell types and tissues
    Article Snippet: .. Vectors were purified using the QIAGEN Plasmid Plus 96 Miniprep Kit (QIAGEN). .. HeLa-S3 cells (JCRB Cell Bank) were cultured in MEM (WAKO) supplemented with 10% FBS (NICHIREI BIOSCIENCE INC., Lot No. 7G0031), 100 Units/mL penicillin and 100 μg/mL streptomycin (both Life Technologies).

    Article Title: Identification of fasciclin-like arabinogalactan proteins in textile hemp (Cannabis sativa L.): in silico analyses and gene expression patterns in different tissues
    Article Snippet: .. Plasmids were extracted using the QIAGEN plasmid miniprep kit and sequenced on an Applied Biosystems 3500 Genetic Analyser using the BigDye Terminator v3.1 Cycle Sequencing and the BigDye XTerminator Purification kits, according to the manufacturer’s instructions. ..

    Article Title: Capped antigenomic RNA transcript facilitates rescue of a plant rhabdovirus
    Article Snippet: .. General Plasmid DNA was extracted using an AxyPrep plasmid miniprep kit (Axygen, Union, CA) or a QIAGEN plasmid miniprep kit (QIAGEN, Hilden, Germany) and used for cloning and sequencing. .. DNAs for subcloning were recovered from gel slices using an AxyPrep DNA gel extraction kit (Axygen, Union, CA) or a Purelink Quick gel extraction kit (Invitrogen, Shanghai, China).

    Article Title: Novel Biocatalysts Based on S-Layer Self-Assembly of Geobacillus Stearothermophilus NRS 2004/3a: A Nanobiotechnological Approach
    Article Snippet: .. Plasmid DNA from transformed cells was isolated with the Qiagen Plasmid miniprep kit. ..

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: .. 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions. .. Measure DNA concentration by a high-throughput NanoDrop 8000 spectrophotometer.

    Article Title: Functional and phylogenetic characterization of noncanonical vitamin B12–binding proteins in zebrafish suggests involvement in cobalamin transport
    Article Snippet: .. Potential destination clones were isolated according to the protocol of the Qiagen Plasmid Miniprep kit (Qiagen) for further use. .. Chinese Hamster Ovarian K1 cells were obtained from liquid nitrogen stocks, thawed, and plated in liquid media consisting of DMEM supplemented with 10% FBS (Thermo Fisher, lot no.: 1901903), 1× Non-Essential Amino Acids (Thermo Fisher), and 2 m m l -glutamine (Thermo Fisher).

    Negative Control:

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: CRITICAL STEP Wells G12 and H12 are used as negative control (water as template) and positive control (10 ng of empty pDEST-AD), respectively. .. 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions.

    Selection:

    Article Title: A Genome-Scale Vector Resource Enables High-Throughput Reverse Genetic Screening in a Malaria Parasite
    Article Snippet: After two rounds of antibiotic selection, cultures were reinoculated into fresh selective medium and incubated for no longer than 16 hr at 37°C to favor the correct recombination product. .. Intermediate vector DNA was obtained using a QIAGEN Plasmid Plus 96 Miniprep kit and eluted into 65 μl Tris-EDTA buffer.

    Article Title: Streambed microstructure predicts evolution of development and life history mode in the plethodontid salamander Eurycea tynerensis
    Article Snippet: Transformed cells were plated on kanamycin/X-gal plates for blue/white selection. .. Plasmids were purified using a Qiagen Plasmid Miniprep kit and standard protocols, except that a 1:10 dilution of EB buffer:/water was used for the final elution.

    Agarose Gel Electrophoresis:

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: TROUBLESHOOTING 16 If the PCR is successful as determined by running a 0.8% agarose gel to observe the expected DNA sizes, prepare glycerol stock plates by combining 80 µl of sterile 40% w/v glycerol with 80 µl of bacterial culture in 96-well round bottom plates. .. 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions.

    Spectrophotometry:

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions. .. Measure DNA concentration by a high-throughput NanoDrop 8000 spectrophotometer.

    Concentration Assay:

    Article Title: Effect of Mutations of Arginine 94 on Proton Pumping, Electron Transfer, and Superoxide Anion Generation in Cytochrome b of the bc1 Complex from Rhodobacter sphaeroides *
    Article Snippet: The expression plasmid pRKD fbc Fm BCH Q was purified from an aliquot of semi-aerobically grown culture using the Qiagen plasmid miniprep kit. .. Because R. sphaeroides cells contain four types of endogenous plasmids, the isolated plasmids lacked the purity and concentration needed for direct sequencing.

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions. .. Measure DNA concentration by a high-throughput NanoDrop 8000 spectrophotometer.

    High Throughput Screening Assay:

    Article Title: Base-resolution stratification of cancer mutations using functional variomics
    Article Snippet: 17 For the remaining bacterial culture in 96-well deep-well plates, perform miniprep plasmid isolation using a QIAgen 96-well plasmid extraction kit, using a bench vacuum setting or a QIAgen liquid handling robot, according to manufacturer’s instructions. .. Measure DNA concentration by a high-throughput NanoDrop 8000 spectrophotometer.

    Gel Extraction:

    Article Title: Capped antigenomic RNA transcript facilitates rescue of a plant rhabdovirus
    Article Snippet: General Plasmid DNA was extracted using an AxyPrep plasmid miniprep kit (Axygen, Union, CA) or a QIAGEN plasmid miniprep kit (QIAGEN, Hilden, Germany) and used for cloning and sequencing. .. DNAs for subcloning were recovered from gel slices using an AxyPrep DNA gel extraction kit (Axygen, Union, CA) or a Purelink Quick gel extraction kit (Invitrogen, Shanghai, China).

    Article Title: Novel Biocatalysts Based on S-Layer Self-Assembly of Geobacillus Stearothermophilus NRS 2004/3a: A Nanobiotechnological Approach
    Article Snippet: Qiagen MinElute gel extraction kit was used to purify DNA fragments from agarose gels and Qiagen MinElute reaction cleanup kit was used to purify digested oligonucleotides and plasmids. .. Plasmid DNA from transformed cells was isolated with the Qiagen Plasmid miniprep kit.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 95
    Qiagen miniprep kits
    Different RNase concentrations do not reduce DNA yield in Miraprepped samples and Miraprep is not significantly contaminated by low molecular weight RNA. (A) Standard <t>Miniprep,</t> or Miraprepped plasmids prepared using 1x volume of ethanol, were treated with indicated RNase concentration, added freshly into the resuspension buffer before beginning the procedure. Top: 0.4 μg was electrophoresed on an agarose gel. DNA concentration only varied slightly when RNase was freshly added. Bottom: OD260/280 ratio. (B,C) Testing for low molecular weight RNA in Miniprep and Miraprep samples, respectively. (B) Miraprep and Miniprep samples of the 8 kb plasmid contain little or no small molecular weight RNA. Pre-column = after alkaline lysis, Flow-through = flow-through of spin column, Final lane in each set is eluted plasmid. 10 μl of pre-column and flow-through samples were loaded, while 2 μl were loaded of Miniprep or Miraprep samples. (C) Miniprep and Miraprep samples of the 14 kb plasmid have little to no low molecular RNA present. Loading same as described in (B).
    Miniprep Kits, supplied by Qiagen, used in various techniques. Bioz Stars score: 95/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/miniprep kits/product/Qiagen
    Average 95 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    miniprep kits - by Bioz Stars, 2020-04
    95/100 stars
      Buy from Supplier

    Image Search Results


    Different RNase concentrations do not reduce DNA yield in Miraprepped samples and Miraprep is not significantly contaminated by low molecular weight RNA. (A) Standard Miniprep, or Miraprepped plasmids prepared using 1x volume of ethanol, were treated with indicated RNase concentration, added freshly into the resuspension buffer before beginning the procedure. Top: 0.4 μg was electrophoresed on an agarose gel. DNA concentration only varied slightly when RNase was freshly added. Bottom: OD260/280 ratio. (B,C) Testing for low molecular weight RNA in Miniprep and Miraprep samples, respectively. (B) Miraprep and Miniprep samples of the 8 kb plasmid contain little or no small molecular weight RNA. Pre-column = after alkaline lysis, Flow-through = flow-through of spin column, Final lane in each set is eluted plasmid. 10 μl of pre-column and flow-through samples were loaded, while 2 μl were loaded of Miniprep or Miraprep samples. (C) Miniprep and Miraprep samples of the 14 kb plasmid have little to no low molecular RNA present. Loading same as described in (B).

    Journal: PLoS ONE

    Article Title: The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields

    doi: 10.1371/journal.pone.0160509

    Figure Lengend Snippet: Different RNase concentrations do not reduce DNA yield in Miraprepped samples and Miraprep is not significantly contaminated by low molecular weight RNA. (A) Standard Miniprep, or Miraprepped plasmids prepared using 1x volume of ethanol, were treated with indicated RNase concentration, added freshly into the resuspension buffer before beginning the procedure. Top: 0.4 μg was electrophoresed on an agarose gel. DNA concentration only varied slightly when RNase was freshly added. Bottom: OD260/280 ratio. (B,C) Testing for low molecular weight RNA in Miniprep and Miraprep samples, respectively. (B) Miraprep and Miniprep samples of the 8 kb plasmid contain little or no small molecular weight RNA. Pre-column = after alkaline lysis, Flow-through = flow-through of spin column, Final lane in each set is eluted plasmid. 10 μl of pre-column and flow-through samples were loaded, while 2 μl were loaded of Miniprep or Miraprep samples. (C) Miniprep and Miraprep samples of the 14 kb plasmid have little to no low molecular RNA present. Loading same as described in (B).

    Article Snippet: Consistent with this, when we compared older Miniprep kits, in which RNase was added to the resuspension buffer several weeks earlier, with new Miniprep kits, where the manufacturer’s RNase was freshly added, DNA yields as assessed by OD260 sometimes were higher than those determined by comparison to DNA markers of known concentrations analyzed via agarose gel electrophoresis (data not shown).

    Techniques: Molecular Weight, Concentration Assay, Agarose Gel Electrophoresis, Plasmid Preparation, Alkaline Lysis, Flow Cytometry

    Miraprepped plasmids can be effectively used to transfect human tissue culture cells. (A) Immunofluorescence of human SW480 cells transfected with a plasmid encoding GFP (3 kb; Miraprepped using 1x volume of ethanol+50 μg/ml RNase) and stained for β-catenin via antibody. SW480 cells have high levels of the Wnt transcriptional co-activator β-catenin due to a mutation in one of its key negative regulators, APC. (A’) GFP is uniformly distributed throughout transfected cells. (A”) Expression of GFP does not alter β-catenin levels—arrows compare a transfected and an untransfected cell. (B) Immunofluorescence of SW480 cells transfected with a plasmid encoding GFP-tagged Drosophila APC2 (8 kb; Miraprep using 1x volume of ethanol+50 μg/ml RNase). (B’) APC2 is uniformly distributed in the cytoplasm. (B”) Fly APC2 is able to reduce β-catenin levels, thus compensating for the mutation of the endogenous human APC in the SW480 cells. (C) Transfection efficiency into SW480 cells is similar for Miraprepped samples (using 1x volume of ethanol+50 μg/ml RNase) and those transfected with DNA prepared via the standard Qiagen Maxiprep procedure. 2 μg of plasmids encoding GFP (3 kb) or GFP-tagged Drosophila APC2 (8 kb) were transfected using Lipofectamine 2000. 100 cells were counted in each of three independent experiments. (D) Immunoblot analysis of transfection efficiency. 3 conditions were tested: DNAs prepared by Miniprep (GeneJET), Miraprep (using 1x volume ethanol), and Miraprep (using 1x vol+50 μg/ml RNase). All led to roughly equal levels of protein expression. Lipofectamine 2000 and 2 μg of plasmid DNA were used. Cells were directly lysed in SDS-loading buffer. aPKCγ was used as the loading control.

    Journal: PLoS ONE

    Article Title: The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields

    doi: 10.1371/journal.pone.0160509

    Figure Lengend Snippet: Miraprepped plasmids can be effectively used to transfect human tissue culture cells. (A) Immunofluorescence of human SW480 cells transfected with a plasmid encoding GFP (3 kb; Miraprepped using 1x volume of ethanol+50 μg/ml RNase) and stained for β-catenin via antibody. SW480 cells have high levels of the Wnt transcriptional co-activator β-catenin due to a mutation in one of its key negative regulators, APC. (A’) GFP is uniformly distributed throughout transfected cells. (A”) Expression of GFP does not alter β-catenin levels—arrows compare a transfected and an untransfected cell. (B) Immunofluorescence of SW480 cells transfected with a plasmid encoding GFP-tagged Drosophila APC2 (8 kb; Miraprep using 1x volume of ethanol+50 μg/ml RNase). (B’) APC2 is uniformly distributed in the cytoplasm. (B”) Fly APC2 is able to reduce β-catenin levels, thus compensating for the mutation of the endogenous human APC in the SW480 cells. (C) Transfection efficiency into SW480 cells is similar for Miraprepped samples (using 1x volume of ethanol+50 μg/ml RNase) and those transfected with DNA prepared via the standard Qiagen Maxiprep procedure. 2 μg of plasmids encoding GFP (3 kb) or GFP-tagged Drosophila APC2 (8 kb) were transfected using Lipofectamine 2000. 100 cells were counted in each of three independent experiments. (D) Immunoblot analysis of transfection efficiency. 3 conditions were tested: DNAs prepared by Miniprep (GeneJET), Miraprep (using 1x volume ethanol), and Miraprep (using 1x vol+50 μg/ml RNase). All led to roughly equal levels of protein expression. Lipofectamine 2000 and 2 μg of plasmid DNA were used. Cells were directly lysed in SDS-loading buffer. aPKCγ was used as the loading control.

    Article Snippet: Consistent with this, when we compared older Miniprep kits, in which RNase was added to the resuspension buffer several weeks earlier, with new Miniprep kits, where the manufacturer’s RNase was freshly added, DNA yields as assessed by OD260 sometimes were higher than those determined by comparison to DNA markers of known concentrations analyzed via agarose gel electrophoresis (data not shown).

    Techniques: Immunofluorescence, Transfection, Plasmid Preparation, Staining, Mutagenesis, Expressing

    Testing whether the columns might act as a filter and verifying DNA yields using comparison to known DNA standards. (A) To determine whether silica spin columns might capture DNA by acting as filters, the Miraprep procedure was followed through the neutralization step with 1x volume of ethanol added or no ethanol added as a control, and then the sample was passed over a simple centrifugal filter (pore size 0.22 μm), the filter was washed following the Miraprep protocol, DNA was eluted from the top surface of the filter, and electrophoresed on an agarose gel. DNA was only recovered after ethanol addition. (B) Silica columns are more efficient than centrifugal filters in capturing plasmid DNA. Comparison of DNA yields using silica columns or centrifugal filter columns; from three independently conducted Mirapreps. (C,D) Standard Miniprep plasmids, or Miraprep plasmids prepared using our final protocol using 1x volume of ethanol, were electrophoresed on an agarose gel and amounts compared to known DNA standards (Thermo Scientific GeneRuler 1 kb DNA Ladder #SM0312 (0.5 μg/μl)). 2 μl DNA plus 5 μl loading buffer were loaded in each lane. (C) GenElute kit. (D) GeneJET kit. Above each gel is the DNA amount calculated from OD260 and below the gel estimates from comparison to DNA markers of known amounts. Image J was used to quantify DNA band intensities in (C,D). DNA amounts calculated by both methods were comparable.

    Journal: PLoS ONE

    Article Title: The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields

    doi: 10.1371/journal.pone.0160509

    Figure Lengend Snippet: Testing whether the columns might act as a filter and verifying DNA yields using comparison to known DNA standards. (A) To determine whether silica spin columns might capture DNA by acting as filters, the Miraprep procedure was followed through the neutralization step with 1x volume of ethanol added or no ethanol added as a control, and then the sample was passed over a simple centrifugal filter (pore size 0.22 μm), the filter was washed following the Miraprep protocol, DNA was eluted from the top surface of the filter, and electrophoresed on an agarose gel. DNA was only recovered after ethanol addition. (B) Silica columns are more efficient than centrifugal filters in capturing plasmid DNA. Comparison of DNA yields using silica columns or centrifugal filter columns; from three independently conducted Mirapreps. (C,D) Standard Miniprep plasmids, or Miraprep plasmids prepared using our final protocol using 1x volume of ethanol, were electrophoresed on an agarose gel and amounts compared to known DNA standards (Thermo Scientific GeneRuler 1 kb DNA Ladder #SM0312 (0.5 μg/μl)). 2 μl DNA plus 5 μl loading buffer were loaded in each lane. (C) GenElute kit. (D) GeneJET kit. Above each gel is the DNA amount calculated from OD260 and below the gel estimates from comparison to DNA markers of known amounts. Image J was used to quantify DNA band intensities in (C,D). DNA amounts calculated by both methods were comparable.

    Article Snippet: Consistent with this, when we compared older Miniprep kits, in which RNase was added to the resuspension buffer several weeks earlier, with new Miniprep kits, where the manufacturer’s RNase was freshly added, DNA yields as assessed by OD260 sometimes were higher than those determined by comparison to DNA markers of known concentrations analyzed via agarose gel electrophoresis (data not shown).

    Techniques: Activated Clotting Time Assay, Neutralization, Agarose Gel Electrophoresis, Plasmid Preparation

    Addition of Ethanol leads to increased plasmid DNA yield. (A) DNA plasmid preps of the indicated plasmids with different concentrations of ethanol. Top: DNA concentration as assessed by OD260, middle: 2 μl of each sample was electrophoresed on an agarose gel and visualized by ethidium bromide staining, bottom: OD260/280 ratio. The GeneJET Plasmid Miniprep kit was used. (B) As in (A) but the Qiagen Miniprep kit was used. (C) Plasmid preparations with the GenElute kit, comparing either the standard Miniprep procedure or the Miraprep (using 1x volume of ethanol). The Mirapreps in (C) included fresh addition of RNase (50 μg/ml) as in the final Miraprep protocol, and values are the average of two experiments, showing mean and standard deviation.

    Journal: PLoS ONE

    Article Title: The Miraprep: A Protocol that Uses a Miniprep Kit and Provides Maxiprep Yields

    doi: 10.1371/journal.pone.0160509

    Figure Lengend Snippet: Addition of Ethanol leads to increased plasmid DNA yield. (A) DNA plasmid preps of the indicated plasmids with different concentrations of ethanol. Top: DNA concentration as assessed by OD260, middle: 2 μl of each sample was electrophoresed on an agarose gel and visualized by ethidium bromide staining, bottom: OD260/280 ratio. The GeneJET Plasmid Miniprep kit was used. (B) As in (A) but the Qiagen Miniprep kit was used. (C) Plasmid preparations with the GenElute kit, comparing either the standard Miniprep procedure or the Miraprep (using 1x volume of ethanol). The Mirapreps in (C) included fresh addition of RNase (50 μg/ml) as in the final Miraprep protocol, and values are the average of two experiments, showing mean and standard deviation.

    Article Snippet: Consistent with this, when we compared older Miniprep kits, in which RNase was added to the resuspension buffer several weeks earlier, with new Miniprep kits, where the manufacturer’s RNase was freshly added, DNA yields as assessed by OD260 sometimes were higher than those determined by comparison to DNA markers of known concentrations analyzed via agarose gel electrophoresis (data not shown).

    Techniques: Plasmid Preparation, Concentration Assay, Agarose Gel Electrophoresis, Staining, Standard Deviation

    Confirmation of HGT from replicating and homology plasmids. ( a ) plasmid DNA isolated with a standard miniprep kit and digested using SspI. Lanes 1–3: Kanamycin-resistant Acinetobacter clones resulting from HGT of pBAV1k. Lanes 4–6: Kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03+homology. Lane 7: E. coli carrying pBAV1k. ( b ) inverse PCR with primers located in the kan gene using genomic DNA isolated from kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03H.

    Journal: eLife

    Article Title: Inter-species population dynamics enhance microbial horizontal gene transfer and spread of antibiotic resistance

    doi: 10.7554/eLife.25950

    Figure Lengend Snippet: Confirmation of HGT from replicating and homology plasmids. ( a ) plasmid DNA isolated with a standard miniprep kit and digested using SspI. Lanes 1–3: Kanamycin-resistant Acinetobacter clones resulting from HGT of pBAV1k. Lanes 4–6: Kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03+homology. Lane 7: E. coli carrying pBAV1k. ( b ) inverse PCR with primers located in the kan gene using genomic DNA isolated from kanamycin-resistant Acinetobacter clones resulting from HGT of pRC03H.

    Article Snippet: We isolated any plasmid DNA from these clones and from the bait E. coli carrying pBAV1k using a standard miniprep kit (Qiagen, Hilden, Germany), digested with SspI, which should cut twice, and ran the result on a gel ( ).

    Techniques: Plasmid Preparation, Isolation, Clone Assay, Inverse PCR