Navigation Links
High Electrotransformation Efficiencies Obtained With DNA From Ligation Mixtures

Paul Zoller, Research and Development Department, Genetic Systems Division, Bio-Rad Laboratories, 2000 Alfred Nobel Drive, Hercules, CA 94547


Introduction
It has become clear that electroporation of E. coli results in significantly higher transformation efficiencies than chemical transformation procedures by a factor of 10 to 100, and is therefore the method of choice for applications requiring high efficiencies. For example, preparation of cDNA libraries from a small amount of plasmid DNA necessitates the high transformation efficiencies obtained using electroporation. Problems in obtaining high efficiencies occur when electroporating DNA directly from ligation mixtures, due to components in the mixtures which interfere with transformation.1 Several approaches have been suggested that may overcome this interference problem, such as dilution, precipitation, microdialysis, and heat treatment of the ligation reaction.2-5 In this study I explored combinations of these approaches to determine the method that results in the highest electro-transformation efficiencies for the introduction of DNA directly from ligation reactions.


Methods
Sample Preparation
Electro-Competent E. coli JS5 cells and supercoiled pUC18 plasmid DNA (10 pg/l) were obtained from Bio-Rad Laboratories. Linear pUC18 plasmid was prepared by digestion of the supercoiled form with EcoRI restriction enzyme (New England Biolabs), followed by extraction with phenol/chloroform, ethanol precipitation, and resuspension in TE buffer (10 mM Tris, pH 8.0, 1 mM EDTA) to a concentration of 10 ng/l. All ligation reactions consisted of DNA
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Electrotransformation of E. coli With M13 DNA
2. Highest Possible Transformation Efficiencies for High-Throughput Applications
3. New Competent Cells for Highest Transformation Efficiencies
4. Molecular Biology:Easy Dephosphorylation and Rapid DNA Ligation
5. Cloning Based on Efficient Three-Fragment Assembly DNA Ligation
6. Structural Analysis of Glycosylated Peptides in Complex Mixtures with Ion Trap MSn
7. LC-MALDI and MALDI-MS/MS for the Analysis of Complex Peptide Mixtures
Post Your Comments:
(Date:12/24/2014)... LOUIS , Dec. 23, 2014 ... today that the waiting period under the Hart-Scott-Rodino ... the acquisition expired on December 22, 2014, thereby ... review requirement for the acquisition of the Company ... U.S. antitrust clearance satisfies another condition to closing ...
(Date:12/22/2014)... Brussels (PRWEB) December 22, 2014 The ... the global leader of technical training across the life ... Clinical Data Management (SCDM) to provide the organization's ... certification programs —providing access to the more than 350 ... SCDM members with 10% off when registering for a ...
(Date:12/22/2014)... , Dec. 22, 2014  Alternative Energy ... that it has signed a letter of intent ... has developed and patented a nanotechnology-based development platform ... products that enable rapid on-site collection and testing ... and health issues in an immediate, non-invasive and ...
(Date:12/22/2014)... The American Journal of ... research, reviews and editorials addressing developments and pathways ... published a provocative article exploring the role of ... potential treatment of prostate cancer. , The ... the possibility that there could be a connection ...
Breaking Biology Technology:Acquisition of Sigma-Aldrich by Merck KGaA, Darmstadt, Germany, Receives U.S. Antitrust Clearance 2Acquisition of Sigma-Aldrich by Merck KGaA, Darmstadt, Germany, Receives U.S. Antitrust Clearance 3CfPIE Announces Partnership with the Society for Clinical Data Management 2CfPIE Announces Partnership with the Society for Clinical Data Management 3ALNE Announces Intention To Acquire BioTechPharma 2Prostate Cancer Experts Dissect the Role of Follicle-stimulating Hormone in the Development, Progression and Potential Treatment of Prostate Cancer 2Prostate Cancer Experts Dissect the Role of Follicle-stimulating Hormone in the Development, Progression and Potential Treatment of Prostate Cancer 3Prostate Cancer Experts Dissect the Role of Follicle-stimulating Hormone in the Development, Progression and Potential Treatment of Prostate Cancer 4
... DIEGO, Feb. 26 Bio-Matrix Scientific Group, Inc. ... today that the San Diego-based biotechnology company is ... Pick" on WallStreetCorner.com ( http://wallstreetcorner.com/stockpick.html ), an editorial ... and Special Situation profiles are regularly read by ...
... for Biomedical,Research today presented its strategies for drug ... A large computational,project with basic research support and ... under the broad-based Epistemic Drug,Discovery(R) project. , ... Institute, Dr. J.R. Prous, explained that "We,are committed ...
... February 26 ScreenCancer AS, a Norwegian,company ... insurance,companies and individuals, today announced that it ... screening test to its European Cancer,Screening Program. ... a subsidiary of Quest Diagnostics Incorporated (NYSE: ...
Cached Biology Technology:Bio-Matrix Scientific Group Featured as 'Stock Pick' in Oakley's Column on WallStreetCorner.com 2Prous Institute Presents Innovative Approach to Drug Discovery 2ScreenCancer Adds InSure(R) FIT(TM) Colorectal Cancer Screening Test to European Cancer Screening Program 2ScreenCancer Adds InSure(R) FIT(TM) Colorectal Cancer Screening Test to European Cancer Screening Program 3
(Date:12/17/2014)... NEW YORK , Dec. 15, 2014 /PRNewswire-USNewswire/ ... a Good Clinical Practices (GCP) audit to confirm ... Administration (FDA) regulations. This accomplishment enables HITLAB to ... trials utilizing the highest principles for patient safety ... to improve global healthcare access, quality, and delivery ...
(Date:12/11/2014)... --Research and Markets , ... ) has announced the addition of the ... to their offering. One major ... technology. With continuous advances in technology, it is ... standard that meets the needs and expectations of ...
(Date:12/10/2014)... RALEIGH, N.C. , Dec. 9, 2014  Valencell, ... announced it is seeing a staggering demand from its ... demand, the Company revealed, is not solely coming from ... industries as well. "A wearable is only ... a result, accuracy is the ultimate driver in long-term ...
Breaking Biology News(10 mins):HITLAB Completes GCP Audit for FDA Smart Device & App Compliance 2Biometrics Market in the APAC Region 2015-2019: Key Vendors are 3M Cogent, NEC, Safran and Suprema 2Wearable Technology Products Demand Highly Accurate Biometric Technology 2
... Sainsbury Laboratory and the John Innes Centre will publish the first ... of disease. They will release the data via a ... system, called GitHub, was also used to crowdsource expertise during the ... around the world will be able to access the RNA sequence ...
... of Illinois at Urbana-Champaign has received a fiveyear, $5.7 million ... corn. These strains have the potential to combat the losses ... A team at the Institute for Genomic Biology in the ... the research. "Ozone can cause major damage and ...
... Since Edison,s first bulb, heat has been a mostly undesirable ... light into heat at the point of need, on the ... The method created by the Rice labs of Michael Wong, ... American Chemical Society journal ACS Nano makes use ...
Cached Biology News:Crowdsourcing to kickstart comeback from ash dieback 2University of Illinois receives grant to study ozone resistance in corn 2Rice uses light to remotely trigger biochemical reactions 2Rice uses light to remotely trigger biochemical reactions 3Rice uses light to remotely trigger biochemical reactions 4
MAGP-2 (C-19)...
Laminin gamma-2 (G-16)...
... Proliferation Kit, For 100 slides. Immunocytochemistry.Sample ... in vitro tissue labeling.Recommended cell number: ... (labeling 1-2 hours).Store at 2-8 C ... Category: Drug Screening & Cellular Assays, ...
Dysadherin 2 (L-15)...
Biology Products: