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:5/23/2017)... ... May 23, 2017 , ... Energetiq Technology, a world ... facility expansion to accommodate its rapid growth. , The renovations at the company’s ... of the existing areas. The expansion includes, a state-of-the-art engineering facility, and a ...
(Date:5/23/2017)... ... May 23, 2017 , ... Customers often prefer PLC ... and again. METTLER TOLEDO has released two new videos that show how they ... of the ACT350 into Siemens and Allen Bradley PLCs is easy and fast. ...
(Date:5/21/2017)... , ... May 19, 2017 , ... ... and educational conference of the American Association of Bioanalysts (AAB) and the College ... in Houston. The conference reinforces AAB’s commitment to excellence in clinical laboratory services ...
(Date:5/18/2017)... Mass. (PRWEB) , ... May 18, 2017 , ... ... the procedure on April 28, 2017 at the Prince Of Wales Private Hospital. ... cervical disc at level C6-C7. The patient failed conservative treatments prior to undergoing ...
Breaking Biology Technology:
(Date:4/3/2017)... April 3, 2017  Data captured by ... platform, detected a statistically significant association between ... to treatment and objective response of cancer ... to predict whether cancer patients will respond ... as well as to improve both pre-infusion potency ...
(Date:3/30/2017)... The research team of The Hong Kong Polytechnic University (PolyU) ... ground breaking 3D fingerprint minutiae recovery and matching technology, pushing contactless ... use in identification, crime investigation, immigration control, security of access and ... ... A research team led by Dr Ajay Kumar ...
(Date:3/29/2017)... 29, 2017  higi, the health IT company that ... North America , today announced a Series B ... of EveryMove. The new investment and acquisition accelerates higi,s ... to transform population health activities through the collection and ... higi collects and secures data today on behalf ...
Breaking Biology News(10 mins):