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:10/11/2017)... HILLS, Calif. , Oct. 11, 2017  SkylineDx today ... (ICR) and University of Leeds ... risk-stratify patients with multiple myeloma (MM), in a multi-centric Phase ... University of Leeds is the sponsor ... and ICR will perform the testing services to include high-risk ...
(Date:10/11/2017)... ... October 11, 2017 , ... A new study published in ... and fresh in vitro fertilization (IVF) transfer cycles. The multi-center matched ... , After comparing the results from the fresh and frozen transfer cohorts, the ...
(Date:10/10/2017)... ... October 10, 2017 , ... San Diego-based team building and cooking events company, ... today. The bold new look is part of a transformation to increase awareness, ... significant growth period. , It will also expand its service offering from its signature ...
(Date:10/10/2017)... , ... October 10, 2017 , ... ... Center’s FirstHand program has won a US2020 STEM Mentoring Award. Representatives of the ... for Excellence in Volunteer Experience from US2020. , US2020’s mission is to change ...
Breaking Biology Technology:
(Date:10/4/2017)... -- GCE Solutions, a global clinical research organization (CRO), announces the launch ... October 4, 2017. Shadow is designed to assist medical writers and ... of the European Medicines Agency (EMA) in meeting the requirements for ... ... ...
(Date:8/15/2017)... HAMPTON, Va. , Aug. 15 2017   ivWatch LLC ... effectiveness of intravenous (IV) therapy, today announced receipt of its ISO ... (QMS) developed by the International Organization for Standardization (ISO®). ... ivWatch Model 400 Continuous Monitoring device for ... "This ...
(Date:6/30/2017)... 30, 2017 Today, American Trucking Associations ... of face and eye tracking software, became the ... program. "Artificial intelligence and advanced ... monitor a driver,s attentiveness levels while on the ... to detect fatigue and prevent potential accidents, which ...
Breaking Biology News(10 mins):