Navigation Links
Highest Transfection Efficiency of an Endotoxin-Sensitive Mammalian Cell,,,Line

ng the cup, and eluting the purified DNA from the matrix in low-ionic-strength buffer. The resulting purified plasmid DNA is ready for subsequent molecular biology applications.

Four different plasmid purification methods were analyzed. Stratagene's StrataPrep EF plasmid midiprep kit, Qiagen's QIAfilter Plasmid Maxi kit with additional Endotoxin Free Buffer set (AEC: anion exchange chromotography), Promega's Wizard PureFection Plasmid DNA Purification system (MB: magnetic beads), and cesium chloride density gradient centrifugation (CsCl).

Yields and Purity

pCMV b-gal plasmid DNA was isolated from Epicurian Coli XL1-Blue MRF E. coli cells with the StrataPrep kit. Yields of plasmid DNA ranged from 360 g using the MB method to 473 g for CsCl. Yields of the StrataPrep Kit and AEC-purified DNA were 392 g and 433 g, respectively, for an equivalent volume of bacterial culture (Table 1).

The 260-nm to 280-nm optical density (OD) ratios ranged between 1.8 and 1.9 for each plasmid preparation (Table 1), indicating a high level of purity.5 OD values at 240 nm were also measured to detect other contaminants such as EDTA6 and detergents. The 260-nm to 240-nm OD ratios ranged between 1.5 and 1.7, which indicated that the plasmid preparations were essentially devoid of these contaminants6 (Table 1).

Endotoxin units per microgram of plasmid DNA (EU/g) were measured (BioWhittaker, Inc.) using the Kinetic QCL assay. Endotoxin levels for the StrataPrep Kit and AEC-purified DNA were approximately 800 times lower than MB-purified DNA and 3 times lower than CsCl-purified plasmid DNA (Table 1).

Transfection of Endo
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Highest Possible Transformation Efficiencies for High-Throughput Applications
2. New Competent Cells for Highest Transformation Efficiencies
3. Optimizing Transfection Conditions for Studying Signal Transduction Pathways
4. Transfection of Green Fluorescent Protein into Human Adrenalcarcinoma Cells
5. Improve Lipid- or Calcium Phosphate-Mediated Transfection of Human Dermal Fibroblasts
6. Versatile Transfection Reagent Offers Low Toxicity and Consistent Performance
7. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells
8. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection of Mammalian Cells
9. High-Efficiency Transfections Achieved with New Low-Toxicity Reagent
10. Efficient Transfection of Neurospora Crassa
11. Eppendorf Multiporator Transfection Protocols for Eukaryotic Cells

Post Your Comments:
(Date:5/21/2015)... YORK , May 21, 2015  Prima Biomed ... company that is striving to become a leader in ... cancer, recently announced that the final CVac data from ... shown a clear trend for a clinically meaningful improvement ... in second remission patients. In the group ...
(Date:5/21/2015)... W. R. Grace & Co. (NYSE: ... Germany has received good manufacturing practice (GMP) certification ... Pharmaceutical Excipient Council (IPEC) Foundation. , ... FP brand of pharmaceutical grade excipient silica gels ... Bay, Maryland (USA) and Sorocaba, Brazil locations. The ...
(Date:5/21/2015)... , May 20, 2015 Research and Markets ... addition of the "2015 Global Survey on ... offering. The primary goal of this ... cytometry instruments and reagents. Key information the survey ... selection of flow cytometers, predominantly used applications for ...
(Date:5/20/2015)... 20, 2015 Veracyte, Inc . ... demonstrating the ability of the company,s molecular classifier ... other interstitial lung diseases (ILDs) using samples obtained ... to help thousands of patients avoid invasive, risky ... diagnosis – a frequent challenge for physicians and ...
Breaking Biology Technology:Correction Continues after Big Move - BrokerBank Securities, Inc. 2Grace European Facility Receives GMP Excipient Certification for SYLOID® FP Silica Gel 2Global Survey on Flow Cytometry Adoption Trends 2015 2Veracyte Presents Preliminary Data Demonstrating Ability of Molecular Classifier to Improve Non-Surgical IPF Diagnosis Using Bronchoscopy Samples 2Veracyte Presents Preliminary Data Demonstrating Ability of Molecular Classifier to Improve Non-Surgical IPF Diagnosis Using Bronchoscopy Samples 3Veracyte Presents Preliminary Data Demonstrating Ability of Molecular Classifier to Improve Non-Surgical IPF Diagnosis Using Bronchoscopy Samples 4Veracyte Presents Preliminary Data Demonstrating Ability of Molecular Classifier to Improve Non-Surgical IPF Diagnosis Using Bronchoscopy Samples 5
... Third Wave Technologies, Inc ., a manufacturer of molecular diagnostic ... Drug Administration for approval of a new Cystic Fibrosis ... calls InPlex, is a genotyping test to determine Cystic Fibrosis ... and in diagnostic testing in newborns and children. Cystic Fibrosis ...
... and lobbyists from AT&T and BellSouth must ... the band back together and no one can stop them. , ... off the largest telecom merger in U.S. history. I always ... company. They are another step closer to proving I was right ...
... a month ago, and already Microsoft's Windows Vista , the ... bruised by critical media reports about security flaws and other shortcomings. ... the beta version. We posed several questions to Justin Jaeck, vice ... information technologies for Veridian Homes , to get their take ...
Cached Biology Technology:AT&T - BellSouth merger: Revestiture continues 2AT&T - BellSouth merger: Revestiture continues 3AT&T - BellSouth merger: Revestiture continues 4AT&T - BellSouth merger: Revestiture continues 5AT&T - BellSouth merger: Revestiture continues 6Wisconsin firms won't rush to adopt Microsoft Vista 2Wisconsin firms won't rush to adopt Microsoft Vista 3Wisconsin firms won't rush to adopt Microsoft Vista 4
(Date:5/22/2015)... May 22, 2015  NXT-ID, Inc. (NASDAQ: NXTD ) ... will be ending its promotional pricing and making the Wocket ... Laurence Savin , Head of Marketing said, "We,ve extended ... until the end of May. To get your wocket at ... we open the store on June 1, you,ll receive a ...
(Date:5/22/2015)... May 22, 2015 According to ... Solution (2D, 3D, Thermal, Emotion, Forensic), by Software (Middleware, Databases), ... and Utilities, Consumer and Home) - Global Forecast to ... from $2.77 Billion in 2015 to $6.19 Billion by ... Browse 79 market data Tables and   43 Figures ...
(Date:5/21/2015)... According to a new ... Software, Services), by Applications (Surveillance (Airborne, Maritime, Land), ... (Military & Defense & Commercial) - Global Forecast ... is expected to grow from $7252.0 Million in ... Compound Annual Growth Rate (CAGR) of 6.6%. ...
Breaking Biology News(10 mins):NXT-ID, Inc Announces Retail Price For Wocket Smart Wallet And Extends Discounted Promotional Price Till End Of May 2NXT-ID, Inc Announces Retail Price For Wocket Smart Wallet And Extends Discounted Promotional Price Till End Of May 3Facial Recognition Market Worth $6.19 Billion by 2020 2Facial Recognition Market Worth $6.19 Billion by 2020 3Thermal Imaging Market Worth $9,998.9 Million by 2020 2Thermal Imaging Market Worth $9,998.9 Million by 2020 3Thermal Imaging Market Worth $9,998.9 Million by 2020 4
... Arlington, VA, March 18, 2010. How did China ... North America and Europe? A panel of U.S. experts ... comparative findings in a workshop at the National Science Foundation ... the sponsorship of the National Science Foundation and other Federal ...
... BOZEMAN, Mont. -- Bile secretions in the small ... to change their behavior to maximize their chances of ... the Society for General Microbiology during its spring meeting ... the findings could allow us to better protect food ...
... toxicologist and a multidisciplinary team of researchers have ... composed of 60 carbon atoms. The team also ... hold hope for treatment of Parkinson,s disease, Alzheimer,s ... appeared in Toxicology and Applied Pharmacology and represents ...
Cached Biology News:Workshop on R&D for rapid vaccine development and production in Asia/Australia 2Montana State researcher discovers that bile sends mixed signals to E. coli 2Carbon nanostructures -- elixir or poison? 2Carbon nanostructures -- elixir or poison? 3
...
... compact GC-8A is Shimadzu's basic gas chromatograph ... to use, the GC-8A supports a single ... column injection port(s), and analog output. Isothermal ... available. The GC-8A is only 17.3 inches ...
The novel and unique HCTplus delivers unmatched information-rich data-dependent LC/MSn data for todays challenge in proteomics and metabonomics work....
... In the six short years since publication ... John Walkers widely acclaimed Protein Protocols Handbook, ... in protein biochemistry. In recognition of the ... the rapid evolution of new methodologies directed ...
Biology Products: