Navigation Links
Jurkat

Multiporator Transfection Protocol Protocol No. 4308 915.020 11/1999 Cell line Jurkat, T-lymphocyte, human leukemia (suspension cell line) Transfection with Plasmid pEGFP-N1 (in bidistilled H2O) Electroporation buffer Eppendorf Hypoosmolar Electroporation Buffer (PH) Culture medium RPMI 1640 / 10% FCS Cuvette Eppendorf, 2 mm gap width, 400 l Temperature RT (20-25 C) Reference Prof. Ulrich Zimmermann Lehrstuhl fr Biotechnologie Biozentrum Universitt Wrzburg
Am Hubland D-97074 Wrzburg Phone +49 931 888 4508 Fax +49 931 888 4509
e-mail: zimmerma@biozentrum.uni-wuerzburg.de
  1. Harvest the cells in the exponential growth phase and centrifuge them (for 5-10 minutes, 200 x g, at room temperature).
  2. Resuspend the cells in RPMI 1640 / 0.5% FCS, determine the number of cells and centrifuge them (for 5-10 minutes, 200 x g, at room temperature). Remove supernatant.

    Note: The overall incubation time in the Eppendorf Electroporation Buffer must not exceed 30 minutes to guarantee a successful electroporation!

  3. Resuspend the cells in Hypoosmolar Electroporation Buffer. When doing so, set the cell concentration to 1 x 106 cells/ml.
  4. Add and mix plasmid DNA (5-20 g/ml final concentration, in bidistilled H2O).
  5. Transfer 400 l cell suspension into electroporation cuvettes (2 mm gap width). The cell suspension must be free of air bubbles.
  6. Electroporation:

    Mode Eukaryotes Voltage (V) 240 V Time constant (T) 40 s No. of pulses (n) 1
  7. After the pulse, allow the cell suspension to stand in the cuvette for 5-10 minutes at room temperature.
  8. Carefully transfer the cell suspension from the cuvette to 5 ml RPMI 1640 / 10% FCS, and cultivate it in a 60 mm culture dish.
    Note: After pulsing, the cells should be incubated for 2-3 h at 37 C before any centrifugation is performed, to ensure resealing of the membrane.
Detection methods for transfection:
The expression of the plasmid pEGFP-N1 can be detected clearly after 24-48 hours with the aid of FACS analysis or under a fluorescence microscope. Result: Survival rate: 85% Transfection rate: 67% based on the number of surviving cells
57% based on the initial number of cells used for the experiment Results were measured 24 hours after transfection.


'"/>

Source:


Page: All 1 2 3

Related biology technology :

1. Kluyveromyces lactis
2. Lactococcus lactis
3. Lactococcus lactis
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Jurkat

(Date:6/30/2015)... ... June 30, 2015 , ... ... She has accepted the position of vice president of instruction at Chippewa Valley ... , “I greatly appreciated the opportunity to lead SIUE during the past three ...
(Date:6/29/2015)... ... June 29, 2015 , ... ... Under the terms of the agreement, MediVet will acquire the rights to a ... MB-007 is currently in pre-clinical development as a potential next-generation treatment for various ...
(Date:6/29/2015)...  Neuralstem, Inc. (NYSE MKT: CUR), a biopharmaceutical company ... and cell therapy treatments for central nervous system diseases, ... on the NASDAQ Capital Market under the symbol "CUR." ... the NYSE MKT until the market close on July ... expected to commence on July 13, 2015. ...
(Date:6/29/2015)... Elsevier , a world-leading ... services, today announced the highlights of its journal Impact ... Citation Reports® (JCR) published by Thomson Reuters, Elsevier saw ... to 2014, ahead of the aggregate across other journals. ... 62 subject categories, up from 61 in 2013. ...
Breaking Biology Technology:SIUE’s Furst-Bowe Accepts VP of Instruction at CVTC 2SIUE’s Furst-Bowe Accepts VP of Instruction at CVTC 3MediVet Biologics Acquires Exclusive Patent License from Kansas State University 2MediVet Biologics Acquires Exclusive Patent License from Kansas State University 3Neuralstem, Inc. Announces Approval for Listing on the NASDAQ Capital Market 2Neuralstem, Inc. Announces Approval for Listing on the NASDAQ Capital Market 3Elsevier Announces 2014 Citation Impact Highlights 2Elsevier Announces 2014 Citation Impact Highlights 3
... Multiporator / Electroporator 2510 , , , , , , , ... Protocol No. 4308 915.521 02/2002 , , , ... , Microorganism , ... Cell type , Bacteria, gram positive, , ...
... , , , , Multiporator / Electroporator 2510 , , , ... Transformation Protocol , Protocol No. 4308 915.514 04/2002 , ... , , , , ... , Cell type , Bacteria, ...
... , , , , Multiporator / Electroporator 2510 , , , ... Transformation Protocol , Protocol No. 4308 915.511 04/2002 , ... , , , , ... , Cell type , Bacteria, gram ...
Cached Biology Technology:Mycobacterium intracellulare 2Escherichia coli K12 2Escherichia coli K12 3Escherichia coli C600 2Escherichia coli C600 3
(Date:6/30/2015)... , June 30, 2015 To bolster its efforts ... HYPR Corp. announced today the addition of two new team ... as board advisor and David Raviv will act ... underscore HYPR Corp.,s commitment to providing the most secure solutions ... Sirota co-founded Layer 7 Technologies, a provider of security ...
(Date:6/30/2015)... Pa. , June 30, 2015 ... the company as CEO to help further develop Genisphere,s ... wealth of partnering experience, having spent much of the ... including Therakine, Amgen, Baxter Bioscience, and Johnson & Johnson. ... focus on Technology and Biotech Corporate Finance. He graduated ...
(Date:6/29/2015)... 24, 2015 Research and ... "Latin America Biomedical Sensors Market - Growth, Trends ... The Latin America Biomedical Sensors market is estimated ... 2.04% over the period 2014-2020 The near ... the genetic formulation of each individual. These sensors can ...
Breaking Biology News(10 mins):HYPR Corp. Expands Biometric Authentication Team with Enterprise All Stars 2HYPR Corp. Expands Biometric Authentication Team with Enterprise All Stars 3Genisphere Appoints New CEO, Thomas Bliss 2Genisphere Appoints New CEO, Thomas Bliss 3Latin America Biomedical Sensors Market Report 2015-2020 2
... ARBOR, Mich. An early laboratory success is taking ... gland transplants that could one day prevent a currently ... The scientists were able to induce embryonic stem ... hormone essential to maintaining bone density. The laboratory results ...
... ANN ARBOR, Mich.---Brain implants that can more clearly record ... at the University of Michigan. The findings could eventually ... as Parkinson,s disease and paralysis. Neural electrodes must ... When the electrodes are implanted, the brain first reacts ...
... calling on Congress to support the Consistency, Accuracy, Responsibility ... (CARE bill), which would ensure that states set minimum ... bill (H.R. 3652) was introduced Sept. 28 in the ... "The introduction of the CARE bill ...
Cached Biology News:Stem cell success points to way to regenerate parathyroid glands 2A step toward better brain implants using conducting polymer nanotubes 2A step toward better brain implants using conducting polymer nanotubes 3SNM urges Congress to support CARE bill 2
Mouse monoclonal antibody raised against a partial recombinant STAU2. NCBI Entrez Gene ID = STAU2...
...
Mouse polyclonal antibody to NR2F2 - nuclear receptor subfamily 2, group F, member 2...
... Pack provides optimized blocking and wash ... Clarifier: Ready-to-use wash solutions have been ... Coated Slides for Microarrays (Product Code ... Buffer: 1L • Microarray Post Hybridization ...
Biology Products: