Navigation Links
Commercial Plasmid Preparation Methods Yield DNA With Different Transfection Capabilities

Kathleen A. Maguire-Zeiss, Brandon K. Harv e y, Rita E. Giuliano and H o w a rd J. Federo ff, University of Rochester, Rochester, NY, USA


Introduction
Transfection efficiency is dependent upon a number of factors, including method of transfection, cell type, and DNA concentration and purity. Liposome-mediated transfection of mammalian cells has proven to yield higher transfection efficiencies and is more reproducible than other methods (Felgner et al. 1987; Rose et al. 1991). However, we have noted a significant difference in transfection efficiency among cell lines and among diff e rent DNA p reparations with liposomemediated transfections. We have sought to determine which method ensures optimal transfection eff iciencies . We prepared plasmid DNA by four commonly employed methods and measured its transfection efficiency in three different cell lines. DNA prepared using modified alkaline lysis followed by purification on Bio-Rads Quantum Prep DNA binding matrix gave superior transfection efficiencies in all three cell lines.


Method
DNA PREPARATION
DNA from the plasmid pHSVlac (Geschwind et al. 1994) was prepared using four different methods. For each method, pHSVlac DNA was prepared from pellets of overnight cultures (250500 ml, grown with antibiotic) by a modified alkaline lysis procedure with further purification according to each suppliers recommended protocol and buffer composition. Method A was a noncommercial reference method; we used the protocol of Saporito-Irwin et al. (1997), which purifies plasmid DNA by repeated precipitation with ammonium acetate . Method D was the protocol supplied by Bio-Rad with it
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. Performance Comparisons of Commercial RT-PCR Systems
2. Comparison of Commercial Dye Terminator Removal Kits, Rev A
3. Efficiently Insert Unique Restriction Sites into Plasmid Vectors
4. High-Efficiency Site-Directed Mutagenesis of Large Plasmid Vectors
5. Simple Purification of DNA from Plasmid Minipreps, PCR Amplifications and Agarose Gels
6. Generate High-Quality, Directional Plasmid cDNA Libraries
7. New Reporter Plasmids for Studying Interferon-Stimulated Signal Transduction Pathways
8. New Fusion Trans-Activator Plasmids for Studying Signal Transduction Pathways
9. Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit
10. Basic Plasmid DNA Isolation Protocol
11. RNA-free Plasmid DNA Isolation Protocol
Post Your Comments:
(Date:7/31/2014)... Rancho BioSciences , the scientific data ... Suite with tranSMART . Scientists that have ... data in tranSMART to Array Suite for further analysis. ... including the new 1.2 release. Array Suite provides ... the analysis of high dimensional quantification data, including expression, ...
(Date:7/31/2014)... 31, 2014 “Rapid technological advances ... the U.S., increasing unemployment, depressing American consumer spending ... is a Catch 22—we need automated manufacturing if ... fewer jobs,” says economist and corporate relocation expert ... Top 10 Pro-Business States for 2014, co-published with ...
(Date:7/31/2014)... The Kidney Cancer Association (KCA) has established ... magazine, Kidney Cancer News. , Bill Bro, KCA CEO, ... maintained by an international news organization. When we began using ... , This month’s program highlights news of an enzyme that ... a recent study. Kidney Cancer News also presents a report ...
(Date:7/30/2014)... Calif. , July 30, 2014 ... a biopharmaceutical company leading the discovery and development of ... report financial results and highlights for the quarter ended ... the U.S. financial markets close. Regulus ... August 6, 2014, at 5:00 pm Eastern Daylight Time ...
Breaking Biology Technology:Rancho BioSciences, in Partnership with Omicsoft, have Integrated Array Suite with tranSMART 2Rancho BioSciences, in Partnership with Omicsoft, have Integrated Array Suite with tranSMART 3ITRA Global Affiliate’s Pollina Corporate Top 10 Pro-Business States Minimize the Impact of Automation on Middle Class and Unemployment 2ITRA Global Affiliate’s Pollina Corporate Top 10 Pro-Business States Minimize the Impact of Automation on Middle Class and Unemployment 3ITRA Global Affiliate’s Pollina Corporate Top 10 Pro-Business States Minimize the Impact of Automation on Middle Class and Unemployment 4Regulus Announces Timing for Second Quarter 2014 Financial Results Webcast and Conference Call 2Regulus Announces Timing for Second Quarter 2014 Financial Results Webcast and Conference Call 3
... most conductive material, discovered in 2004 at the University ... Novoselov, has the potential to revolutionize material science. , ... scientists the Nobel Prize for Physics last year and ... plans for a 50m graphene research hub to be ...
... Novelda AS announced today two new antennas for its CMOS Impulse ... on a Sinuous Antenna geometry. The antenna itself consists of two ... the antenna itself; the lower PCB is a balancing network. The ... can be lowered by attaching a lens. This lens is built ...
... AlloSource, ( www.allosource.org ) a leading developer, ... use in surgical procedures around the world, is among ... Denver Business Journal,s inaugural awards program. With concerns ... employee benefits, the Denver Business Journal sought ...
Cached Biology Technology:Graphene's 'Big Mac' creates next generation of chips 2Wideband Antenna for Novelda Nanoscale Impulse Radar 2AlloSource Named One of Denver's Healthiest Employers 2AlloSource Named One of Denver's Healthiest Employers 3
(Date:7/31/2014)... Hibiscus flowers- they are an iconic symbol of ... in the landscape. Some, like Hawaii,s State Flower- ... Only a relatively few botanists and Hawaiian conservation ... and intriguing related group of plants known as ... Brother of Hibiscus species are in fact highly ...
(Date:7/31/2014)... NXT-ID, Inc., (NXTD: OTCQB) a biometric authentication company ... and consumers wanting to learn more about its  ... on-air today, July 31 st . ... featured in a segment of NewsWatch, a weekly ... the History Channel, FYI Network today. ...
(Date:7/30/2014)... uses reversible epigenetic processes to regulate its genes, and ... a picture of the genome regulation taking place in ... Genome Biology and illustrates how the truffle ... genes,. The authors say this may shed light on ... Black truffles (Tuber melanosporum), also known as Prigord truffles, ...
Breaking Biology News(10 mins):Brother of Hibiscus is found alive and well on Maui 2NXT-ID's Next Generation Biometric Smart Wallet Featured on NewsWatch Consumer Show on the History Channel and FYI Network Today, July 31st 2NXT-ID's Next Generation Biometric Smart Wallet Featured on NewsWatch Consumer Show on the History Channel and FYI Network Today, July 31st 3How black truffles deal with the jumpers in their genome 2
... tiny luminescent particles made of semiconductors, hold promise ... doctors were to use them in humans, quantum ... and possible toxicity. Scientists at Emory University ... a way around those limitations by exploiting a ...
... Calif.) The apparent success of a case in which ... marrow transplant comes as no surprise to Gerhard Bauer, a ... more than 10 years on a similar cure for AIDS ... patient with stem cells that have been engineered to resist ...
... discovered that management efforts intended to assist migrations of ... fish populations. Juveniles that are transported downstream on boats ... breeding grounds, reducing their survivorship and altering adaptations in ... place for over three decades to improve the survival ...
Cached Biology News:'Strained' quantum dots show new optical properties 2UC Davis researchers exploring gene therapy to fight AIDS 2UC Davis researchers exploring gene therapy to fight AIDS 3Transporting juvenile salmon hinders adult migration 2Transporting juvenile salmon hinders adult migration 3
... Platypus Contact Printing Kit contains choice ... with series of 100 / 200 micron ... or 200 um x 10 mm lines, ... 20, 40 or 60 nm and pitch ...
Request Info...
Sterile glass 5 mm beads that make spreading bacterial and yeast cultures easy. Can be washed, autoclaved and reused....
... are supplied as 10X concentrated solutions. REact Buffer ... of the REact buffers and one 250 microl ... four 1 ml vials of REact 1 Buffer ... four 1 ml vials of REact 3 Buffer ...
Biology Products: