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:3/20/2017)... ... 2017 , ... SSCI and Whitehouse Laboratories, divisions of Albany Molecular Research, Inc, ... by the Parenteral Drug Association (PDA), the New York Interphex Show will open on ... and more than 625 exhibitors, the educational and networking opportunities are extremely valuable. INTERPHEX ...
(Date:3/20/2017)... ... March 20, 2017 , ... ... biotech companies, recently announced it will debut a brand new pressure vessel, the ... intelliVessel is controlled by a touch screen panel and features other revolutionary operations ...
(Date:3/20/2017)... Yorba Linda, CA (PRWEB) , ... March 20, ... ... interference and require expensive and laborious sample preparation methods in order to reduce ... is often underappreciated outside of the area of elemental analysis. , This ...
(Date:3/20/2017)... , ... March 20, 2017 , ... The fourth annual ... while innovating to make an extraordinary impact on peoples’ lives, launches today. Sponsored by ... challenge every 5 weeks and awards thousands in cash prizes to engineers, inventors and ...
Breaking Biology Technology:
(Date:2/24/2017)... , Feb. 24, 2017  EyeLock LLC, a leader ... its elite iris biometric solution on the latest ... LTE at Mobile World Congress 2017 (February ... Booth in Hall 3, Stand 3E10. ... Qualcomm Haven™ security platform—a combination of hardware, ...
(Date:2/14/2017)... N.C. , Feb. 14, 2017  Wake Forest ... M.D., as its new chief executive officer (CEO). Freischlag ... CEO John D. McConnell , M.D., who last ... position at the Medical Center, after leading it since ... the full scope of Wake Forest Baptist,s academic health ...
(Date:2/8/2017)... YORK , Feb. 8, 2017 About ... individual,s voice to match it against a stored ... such as pitch, cadence, and tone are compared ... require minimal hardware installation, as most PCs already ... for different transactions. Voice recognition biometrics are most ...
Breaking Biology News(10 mins):