Navigation Links
Pediococcus spp.

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.540 07/2002 Microorganism Pediococcus spp. Cell type Bacteria, gram positive Molecules injected Plasmid DNA (pGK12 and pPN-1) Growth medium MRS-G with 0.5 M sorbitol, 3 % glycine and 40 mM DL-threonine Washing solution 0.5 M sorbitol, 10 % glycerol Electroporation solution 0.5 M sorbitol, 1 mM K2HPO4, 1mM MgCl2, pH 7.0 Outgrowth medium MRS with 0.5 M sorbitol, 20 mM MgCl2 and 2 mM CaCl2 Cuvette 1 mm gap width Reference Caldwell, S. L. et al 1996 Applied and Environmental Microbiology 62, No. 3 936-941 Making electrocompetent cells:

1. Cultivate cells by adding 12 ml overnight preculture (grown in MRS-G with 0.5 M sorbitol) to 800 ml growth medium. Incubate for 2 to 4 hours at 37 C to a cell density of O.D.600 of 0.4-0.6. 2. Harvest by centrifugation. 3. Wash twice in 25 ml washing solution. 4. Resusp end in 1 ml electroporation solution.

Electroporation of cells:

  1. Add 400-600 ng plasmid DNA to 80 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into cuvette.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 1,800 V Time constant (T) 5 ms
  4. Immediately add 2 ml outgrowth medium and keep on ice for approx. 5 min. Incubate for 2 h at 37 C.
  5. Plate aliquots onto selective agar plates; incubate 2-5 days at 37 C.
Expected Results: Transformation efficiency up to 4.6 x 103 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Pediococcus spp.
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:8/15/2017)... PA (PRWEB) , ... August 15, 2017 , ... ... one of the largest Asian exhibitions for analytical and scientific instruments. This year’s ... will be “New Approaches in Mass Spectrometry for Bioanalytical Applications.” This dynamic presentation ...
(Date:8/15/2017)... ... August 15, 2017 , ... Coffea arabica accounts for 70 ... factors. During this educational webinar, participants will learn about the importance of genomics ... better understanding of how genomics is important for coffee breeding improvement. , Attendees ...
(Date:8/14/2017)... ... August 14, 2017 , ... The Conference Forum ... which will take place on September 6, 2017 at the Marriott Copley Place in ... of Experimental Medicine, Informatics, and Regulatory Strategy, Pfizer Innovative Research Lab, Pfizer, who leads ...
(Date:8/11/2017)... ... August 11, 2017 , ... “There ... particular, more natural alternatives to synthetic ingredients,” said Matt Hundt, President of Third ... the established manufacturing presence and know-how of Biorigin will allow us to bring ...
Breaking Biology Technology:
(Date:4/11/2017)... No two people are believed to ... York University Tandon School of Engineering and Michigan ... partial similarities between prints are common enough that ... and other electronic devices can be more vulnerable ... in the fact that fingerprint-based authentication systems feature ...
(Date:4/5/2017)...  The Allen Institute for Cell Science today announces ... portal and dynamic digital window into the human cell. ... application of deep learning to create predictive models of ... a growing suite of powerful tools. The Allen Cell ... publicly available resources created and shared by the Allen ...
(Date:4/4/2017)... 4, 2017   EyeLock LLC , a leader ... United States Patent and Trademark Office (USPTO) has issued ... linking of an iris image with a face image ... the company,s 45 th issued patent. ... timely given the multi-modal biometric capabilities that have recently ...
Breaking Biology News(10 mins):