Navigation Links
Transient Gene Expression Following Single and Double Exponential Decay Electroporation Pulses

in a 0.2 cm gap electroporation cuvette, then the cell suspension was added. Exponential decay electroporation pulses were administered as specified in Table 1 by a Gene Pulser II system with Capacitance Extender II or an EasyJect Plus electroporator (EquiBio) according to the manufacturers instructions. The cells were then cultured in 4 ml DMEM containing 10% serum in hexwell plates in a humidified 37 C, 5% CO2 environment.

Plasmids and Reporter Enzyme Assays
The expression plasmids pCH1104 and pSVZALAΔ5'5, which contain the SV40 early promoter, were used to produce E. coli β-galactosidase and firefly luciferase respectively. These reporter enzyme activities were assayed in cell extracts prepared 24 hours after electroporation. Adherent cells were washed twice with cold PBS and lysed in 300 l luciferase lysis buffer. Extracts were centrifuged briefly to remove cell debris. Luciferase was quantitated using luciferin (Analytical Luminescence Laboratory, San Diego) employed according to the manufacturers protocol, collecting the light produced by 20l of extract for 10 sec with an MGM Optocomp I luminometer. Galactosidase was determined in the same cell extracts using the chemiluminescent Galacto-Light assay (Tropix, Cambridge, Massachusetts) in the same luminometer.


Conclusion
A single, capacitor discharge electroporation pulse gave better transformation results than a Twin-pulse regime. A single pulse treatment is also much easier to optimize, since there are only two electrical parameters to set.


References
1. EasyJect Electroporator Instruction Manual, Stratagene, La Jolla, Ca
'"/>

Source:


Page: All 1 2 3 4 5

Related biology technology :

1. An Epitope Tagging Vector for Gene Expression in Mammalian Cells
2. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
3. High-Level Expression of Peanut Allergens Affected by Rare Codon Usage
4. Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
5. Innovative Tissue Array Technology for High-Throughput Screening of Gene Expression
6. New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter
7. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
8. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
9. A New Lambda Vector for Mammalian Expression
10. Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
11. Codon Bias-Adjusted BL21 Derivatives for Protein Expression
Post Your Comments:
(Date:7/30/2015)... Dallas, Texas (PRWEB) , ... July 30, 2015 , ... ... professional and in-depth study on the current state of the global Propanol market with ... are Sinopec, CNPC, BASF, Eastman, Dow, Carboclor, Mitsui Chemicals, Oxea, ISU Chemical, Sanofi, Royal ...
(Date:7/30/2015)... --  Senomyx , Inc. (NASDAQ: SNMX ), a ... develop, and commercialize novel flavor ingredients for the food, ... for the second quarter 2015. "Moving ... track to achieve our commercial and financial goals," stated ... Company. "Since our last quarterly earnings report, our two ...
(Date:7/30/2015)... 30, 2015  Discovery Laboratories, Inc. (Nasdaq: ... the second $1.0M tranche under a previously awarded ... valued at up to $3.0 million to support ... as a potential medical countermeasure to mitigate acute ... awarded an initial $1.0 million under this grant ...
(Date:7/29/2015)... 2015 Sanofi, un leader ... résultats pour le premier semestre 2015. Le Directeur ... Visionner l,interview vidéo et lire la ... Au sommaire de l,interview :  ... croissance - Diabète - ...
Breaking Biology Technology:Global Propanol Market Technical Data and Manufacturing Analysis 2020 2Global Propanol Market Technical Data and Manufacturing Analysis 2020 3SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 2SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 3SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 4SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 5SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 6SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 7SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 8SENOMYX ANNOUNCES SECOND QUARTER 2015 FINANCIAL RESULTS 9Discovery Labs Receives $1.0 million under an SBIR Grant in Continuing Support of the Development of Aerosolized KL4 Surfactant to Address Radiation-Induced Lung Injury 2Discovery Labs Receives $1.0 million under an SBIR Grant in Continuing Support of the Development of Aerosolized KL4 Surfactant to Address Radiation-Induced Lung Injury 3
... /PRNewswire/ - DNA Genotek, a leading provider of ... announced that The Anthony Nolan Trust, the UK,s largest ... a pilot project aimed at increasing donor recruitment. Bone ... HLA DNA testing to match leukemia patients with prospective ...
... ... leading technology services firm specializing in the intelligent integration of eCommerce, CRM and business ... to build an eCommerce system that will leverage its five divisions: Training, Field Services, ... ...
... CONVENTION, CHICAGO, Illinois , May 5, 2010 ... treatment for tuberculosis,(TB) will shortly start phase II clinical ... novel therapeutic vaccine called RUTI(R),in conjunction with an antibiotic. ... which reduces side effects and healthcare costs, by preventing,re-infection ...
Cached Biology Technology:DNA Genotek's Oragene(R) DNA Product Selected by The Anthony Nolan Trust for Pilot Project 2DNA Genotek's Oragene(R) DNA Product Selected by The Anthony Nolan Trust for Pilot Project 3DNA Genotek's Oragene(R) DNA Product Selected by The Anthony Nolan Trust for Pilot Project 4EDL Consulting Chosen to Implement Visionary Cloud-based eCommerce-Driven Learning Management System 2EDL Consulting Chosen to Implement Visionary Cloud-based eCommerce-Driven Learning Management System 3Archivel Farma's TB Vaccine May Cut Treatment Time From Nine Months to One 2Archivel Farma's TB Vaccine May Cut Treatment Time From Nine Months to One 3
(Date:6/25/2015)... , June 25, 2015  TAKE Solutions Ltd., a ... by the United States Patent and Trademark Office (USPTO) ... process leverages TAKE Solutions, Clinical Accelerators to reduce the ... (when compared to standardization without the accelerators), thus reducing ... At the heart of the patented ...
(Date:6/24/2015)... SAN JOSE, Calif. , June 24, 2015 ... leading developer of human interface solutions, today announced ... fingerprint sensor to provide secure authentication for its ... Having recently reached more than 200 million shipments ... shipment with Sharp reinforces Synaptics, strength, scalability and ...
(Date:6/23/2015)... June 23, 2015   MedNet Solutions , ... the entire spectrum of clinical research, is pleased ... ™ , the company,s intuitive, flexible and affordable ... Silver 2015 Stevie® Award by the American ... Services Website category.  The American Business Awards are ...
Breaking Biology News(10 mins):TAKE Solutions Awarded Patent By USPTO 2Synaptics Brings Natural ID Technology to Sharp's Newest Flagship Smartphone 2MedNet Solutions' iMedNet eClinical Wins Silver Stevie Award 2
... HOUSTON (Nov. 8, 2012) Four young scientists ... Research Institute (NSBRI). Through this two-year program, they conduct ... during long-duration spaceflights. In addition to receiving mentorship from ... are members of one of NSBRI,s seven research teams. ...
... in removing the extra copy of chromosome 21 in ... In Down syndrome, the body,s cells contain three ... A triplicate of any chromosome is a serious ... one-quarter of pregnancy loss from spontaneous miscarriages, according to ...
... 2012  Genomic Health, Inc. (Nasdaq: GHDX ) today ... Officer, Chairman and President will present at the following conferences ... on Thursday, November 15, 2012 at 2:00 p.m. Mountain Time ... Jaffray Healthcare Conference on Tuesday, November 27 at 2:30 p.m. ...
Cached Biology News:Space research institute awards postdoctoral fellowships to 4 scientists 2Extra chromosome 21 removed from Down syndrome cell line 2Extra chromosome 21 removed from Down syndrome cell line 3Genomic Health to Present at Two Upcoming Investor Conferences 2Genomic Health to Present at Two Upcoming Investor Conferences 3
Recombinant Human Erythropoietin (Tissue Culture Grade)...
One-step, microplate or cuvet, colorimetric, detection range 0.78 ug/dL to 65 ug/dL Zn in serum, plasma, urine, saliva etc. Procedure: 10 min....
Human PDGF R beta MAb (Clone # PR7212)...
Recombinant Mouse IL-23, CF...
Biology Products: