Navigation Links
Azotobacter vinelandii

Multiporator / Electroporator 2510 Transformation Protocol

Protocol No. 4308 915.503 04/2002

Microorganism Azotobacter vinelandii Cell type Bacteria, gram negative Molecules injected Plasmid DNA Growth medium Nitrogen-free medium Washing solution Ice-cold 10% sterile glycerol Electroporation solution Ice-cold 10% sterile glycerol Outgrowth medium Azotobacter growth (AG) medium Cuvette 1 mm gap width Reference Kornyi, P. et al 1998 Research in Microbiology 149 361-372

Making electrocompetent cells:

  1. Cultivate cells with vigorous shaking at 30 C to an O.D.620 of 0.4-0.5.

  2. Harvest by centrifugation at 5,000 rpm for 10 min at 0 C.

  3. Wash with the original culture volume of ice-cold 10% glycerol.

  4. Repeat this step three times using a half and a quarter of the original volume, and finally 2-4 ml of the glycerol solution (7.0 x 107 cells/ml final cell concentration).

  5. Freeze 40 l aliquots in liquid nitrogen and store at 70 C.

Electroporation of cells:

  1. Add 1 l (300 ng/l) plasmid DNA to 40 l of electrocompetent cells. Homogenize by gently mixing with pipette

  2. several times. Transfer mixture into a prechilled cuvette.
  3. Wipe moisture from the cuvette and insert the cuvette into the device.

  4. Electroporation:

  5. Mode Prokaryotes O Voltage (V) 1,800 V Time constant (T)
    5 ms No. of pulses (n) 1
  6. Immediately add 1 ml AG medium and incubate with shaking at 200 rpm for 1 h at 30 C.

  7. Plate onto selective AG media plates.

Expected results:

Transformation efficiency up to 6.8 x 105 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Azotobacter vinelandii
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: Azotobacter vinelandii

(Date:7/29/2015)... , July 29, 2015  Pfenex Inc. (NYSE ... financial results will be released on Thursday, August 13, ... Eastern Time, Pfenex management will host a conference call ... update.  A press release outlining the financial results and ... call. Please call 1-866-376-8058 (US) or 1-412-542-4131 ...
(Date:7/29/2015)... /PRNewswire/ - Aeterna Zentaris Inc. (NASDAQ: AEZS ) ... optimized Erk inhibitor molecule for development, thus achieving ... new class of potential cancer therapies. ... for therapeutic intervention in cancer. Recently approved compounds ... B-Raf and Mek inhibitors. Erk inhibitors may be ...
(Date:7/29/2015)... (PRWEB) , ... July 29, 2015 , ... Costello served ... levee system to protect the Orange County, Texas area from future storm surge ... design team in determining several potential levee alignment alternatives for providing protection, Costello also ...
(Date:7/28/2015)... July 29, 2015 A ... acquiring, rapidly developing and commercialising innovative medicines to ... Highly experienced management team; blue chip ...   Mereo BioPharma Group Ltd ("Mereo"), a recently-formed ... ( c. £76.5m), gross, from blue chip institutional investors ...
Breaking Biology Technology:Pfenex To Report Second Quarter 2015 Results and Provide Business Update on Thursday, August 13, 2015 2Aeterna Zentaris Announces Optimized Erk Inhibitor Compounds For Further Development 2Aeterna Zentaris Announces Optimized Erk Inhibitor Compounds For Further Development 3Preventing Storm Surge Flooding in Houston 2Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 2Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 3Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 4Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 5Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 6Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 7Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 8Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 9Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 10Mereo BioPharma Announces Formal Launch of Company; Raises $119m (c.£76.5m) and Acquires Innovative Mid-Stage Clinical Product Portfolio From Leading Pharmaceutical Company 11
... 8 LS9, Inc., the,Renewable Petroleum Company(TM), today ... Officer. Mr. Haywood brings over 30 years of,experience ... track record of,planning, engineering, execution and start-up of ... year will be pivotal to LS9 as the ...
... Circassia to Develop Dopexamine as Part of Proprietary Post-Surgery ... ... October 8 Circassia Ltd, a specialty,biopharmaceutical company focused on ... has acquired the US, Canadian and Japanese,development and commercialization rights ...
... YORK, Oct. 7 Living Proof, a technology-based,beauty ... introduction, No Frizz -- a line of silicone-free,frizz-eliminating ... the company. The Living Proof team of scientists ... Labs and an Institute Professor at MIT., ...
Cached Biology Technology:LS9, Inc. Appoints Bill Haywood Chief Executive Officer 2Circassia Acquires North American and Japanese Rights to Dopexamine 2Circassia Acquires North American and Japanese Rights to Dopexamine 3Living Proof, Inc.: The Launch of the Next Beauty Revolution 2Living Proof, Inc.: The Launch of the Next Beauty Revolution 3
(Date:7/27/2015)... July 27, 2015   Zynx Health ™, ... clinical improvement solutions, today announced that its ... available on Android smartphones and tablets. With this ... organizations can use ZynxCarebook to securely exchange messages ... streamline care transitions to other care settings, and ...
(Date:7/21/2015)... July 21, 2015 Today, ZTE announced its ... as well as expected revenues in 2015 that relate to sales ... revenue guidance of approximately 2,200 MSEK for 2015. Jörgen ... leading smartphone manufacturer in China and we ... 02 5 for Axon , ...
(Date:7/20/2015)... 20, 2015  Acuity Market Intelligence,s latest research ... of Commerce and Privacy" forecasts that between 2014 ... will be downloaded to smart mobile devices by ... market is projected to generate more than $67.9 ... seven-year forecast period.    "Biometrics is ...
Breaking Biology News(10 mins):Zynx Health Adds Android Device Support to ZynxCarebook Solution 2Zynx Health Adds Android Device Support to ZynxCarebook Solution 3FPC's Touch Fingerprint Sensor FPC1025 in ZTE's Smartphone Axon 2Biometrics on Smart Mobile Devices to Redefine Digital Identity with 12.9 Billion Biometric App Downloads between 2014 and 2020 2
... of Community Health (MDCH) has unveiled the state,s ... significant recommendations developed from medical research conducted by ... Kennedy Shriver National Institute for Child Health and ... the Wayne State University School of Medicine. ...
... bottles from the drugstore the human body makes ... University have found a way to use naturally occurring ... Their method also serves as a highly sensitive hydrogen ... role in cellular health and disease. In a ...
... have known for some time that throwing off the ... fact, workers whose sleep-wake cycles are disrupted by night ... as diabetes, obesity and cancer. Researchers at the ... possible molecular link between circadian rhythm disturbances and an ...
Cached Biology News:State of Michigan adopts NIH's PRB progesterone therapy to combat infant mortality 2State of Michigan adopts NIH's PRB progesterone therapy to combat infant mortality 3Scientists discover molecular link between circadian clock disturbances and inflammatory diseases 2Scientists discover molecular link between circadian clock disturbances and inflammatory diseases 3
... Choice Microsphere/Microarray Stabilizer is a is a ... This product is an aqueous solution ... buffer (phosphate buffered saline), pH 6.6 - ... of 0.02% methylisothiazolone and 0.02% bromonitrodioxane as ...
... F1 antigen, Monoclonal Antibody, Unconjugated ... Use at an assay dependent dilution. ... dilution. WB: Use at an assay ... kDa. Not tested in other applications. ...
... formulated for mounting tissue specimens, cell smears, ... immunohistochemical methods for viewing by light microscopy. ... such as AEC, that are alcohol-soluble or ... when slides are cover-slipped forming a coating ...
... is formulated for covering tissue sections and cell preparations immunohistochemically ... products. , ... Application: Vision ... AEC that require an aqueous mounting medium, or those that ...
Biology Products: