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:11/30/2016)... , Nov. 30, 2016 /PRNewswire/ - Portage Biotech Inc. ... Exchange: PBT.U), is excited to announce the formation ... on developing preclinical ophthalmology assets through proof of ... anti-inflammatory created by Portage Pharmaceuticals Limited and being ... ocular surface and anterior segment diseases. This agent ...
(Date:11/30/2016)...  Tempus, a technology company focused on supporting ... Cancer Center have partnered to better determine which ... treatment based on next generation genomic and transcriptomic ... research collaboration, Tempus will provide sequencing and analysis ... to Penn. Utilizing next-generation sequencing, machine learning and ...
(Date:11/30/2016)... --  Merck , a leading science and technology company, ... of agreements with Evotec AG, whereby Evotec AG will ... such as CRISPR and shRNA libraries. Combining access to ... pathway to explore and identify new drug targets. ... targets, a process that can be time- and labor-intensive," ...
(Date:11/30/2016)... Park, North Carolina (PRWEB) , ... November 30, 2016 , ... ... approved the names and symbols for four elements: nihonium (Nh), moscovium (Mc), tennessine ... a 5-month period of public review, the names earlier proposed by the discoverers have ...
Breaking Biology Technology:
(Date:11/24/2016)... Nov. 23, 2016 Cercacor today introduced Ember ... their trainers non-invasively measure hemoglobin, Oxygen Content, ... and Respiration Rate in approximately 30 seconds. Smaller than ... and immediate access to key data about their bodies ... training regimen. Hemoglobin carries oxygen to ...
(Date:11/21/2016)... VILNIUS, Lithuania , Nov. 21, 2016 /PRNewswire/ ... identification and object recognition technologies, today announced that ... for smart cards was submitted for the ... and successfully passed all the mandatory steps of ... III evaluation is a continuing test of fingerprint ...
(Date:11/17/2016)... 17, 2016  AIC announces that it has just released a new white paper ... high-performance scale-out plus high speed data transfer storage solutions. Photo - ... ... ... Setting up a high performance computing or HPC ...
Breaking Biology News(10 mins):