Navigation Links
Clostridium botulinum

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.509 12/2001 Microorganism Clostridium botulinum Cell type Bacteria, gram positive Molecules injected Plasmid DNA (pGK12 in water) Growth medium TPGY medium Washing solution 10% PEG 8000 Electroporation solution 10% PEG 8000 Outgrowth medium TPGY medium with 25 mM MgCl2 Cuvette 4 mm gap width Reference Zhou Y. and Johnson E. A. 1993 Biotechnology Letters 15, No. 2 121-126 Making electrocompetent cells:

1. Grow cells in TPGY medium at 37 C to an O.D.660 of 0.8. 2. Harvest by centrifugation. Wash in 0.8 volumes of PEG at 4 C, keep cells cold during the entire procedure. 3. Resuspend cells in 0.8 ml 10% PEG (number of cells: 4 x 108 cells/ml), keep on ice.

Electroporation of cells:

  1. Add 1 g plasmid DNA to 800 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into a prechilled cuvette. Keep on ice for 2 min.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 2,500 V Time constant (T) 5 ms
  4. Dilute cells into 10 ml TPGY medium with 25 mM MgCl2 and incubate 5 hours at 34 C.
  5. Plate cells on selective TPGY agar plates; incubate at 34 C for 2-3 days.
Expected Results: Transformation efficiency up to 3 x 103 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Clostridium botulinum
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:2/11/2016)... Feb. 11, 2016  Bioethics International, a not-for-profit organization focused ... developed, marketed and made accessible to patients around the world, ... had named the publication of the Good Pharma Scorecard ... is also featured as one of BMJ Open ,s ... year that are most frequently read. Ed Sucksmith ...
(Date:2/11/2016)... 11, 2016 --> ... "Company") (OTCQB: PSID), a life sciences company focused ... Thermomedics subsidiary, which markets the Caregiver® FDA-cleared non-contact ... in January 2016, including entering into agreements with ... sales growth, and establishing several near-term pipeline opportunities. ...
(Date:2/10/2016)... MONTREAL , Febr. 10, 2016 /PRNewswire/ - BioAmber Inc. ... is pleased to announce that Mitsui & Co. Ltd., ... bio-based succinic acid plant, is investing an additional CDN$25 ... equity, increasing its stake from 30% to 40%.  Mitsui ... of bio-succinic acid produced in Sarnia ...
(Date:2/10/2016)... ... February 10, 2016 , ... ... that it has joined the Human Vaccines Project, a public-private partnership to ... cancer. , The Human Vaccines Project brings together leading pharmaceutical and ...
Breaking Biology Technology:
(Date:1/25/2016)... SEATTLE , Jan. 25, 2016  Glencoe Software, ... biotech, pharma and publication industries, will provide the data ... Phenotypic Screening Centre (NPSC). ... Phenotypic analysis ... even whole organisms, allowing comparisons between states such as ...
(Date:1/20/2016)...   MedNet Solutions , an innovative SaaS-based eClinical ... research, is pleased to announce the attainment of record-setting ... result of the company,s laser focus on (and growing ... it,s comprehensive, easy-to-use and highly affordable cloud-based technology platform. ... MedNet growth achievements in 2015 include: , ...
(Date:1/13/2016)... January 13, 2016 ... addition of the  "India Biometrics Authentication ... Forecast (2015-2020)"  report to their ... has announced the addition of the  ... - Estimation & Forecast (2015-2020)" ...
Breaking Biology News(10 mins):