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:6/23/2016)... , June 23, 2016  Blueprint Bio, a company ... to the medical community, has closed its Series A ... Nunez . "We have received a commitment ... capital we need to meet our current goals," stated ... us the runway to complete validation on the current ...
(Date:6/23/2016)... ... June 23, 2016 , ... Velocity Products, a ... designed, tuned and optimized exclusively for Okuma CNC machining centers at The International ... a collaboration among several companies with expertise in toolholding, cutting tools, machining dynamics ...
(Date:6/22/2016)...  According to Kalorama Information, the dominant trends ... include significant efforts in automation as well as ... affordable sequencers, say the healthcare market research firm, ... sample prep materials.  The healthcare market research company,s ... Next Generation Sequencing (NGS) , highlights major trends ...
(Date:6/22/2016)... ... June 22, 2016 , ... The Immigrant ... achievements and contributions to North Texas and the nation, recently held its annual ... community to the civic and economic vitality of North Texas. Proceeds from the ...
Breaking Biology Technology:
(Date:6/9/2016)... leader in attendance control systems is proud to announce the introduction of fingerprint attendance ... the right employees are actually signing in, and to even control the opening of ... ... ... Photo - ...
(Date:6/2/2016)... NEW YORK , June 2, 2016   The ... (Weather), is announcing Watson Ads, an industry-first capability in which ... advertising, by being able to ask questions via voice or ... Marketers have long ... with the consumer, that can be personal, relevant and valuable; ...
(Date:5/16/2016)... , May 16, 2016   EyeLock LLC , ... announced the opening of an IoT Center of Excellence ... and expand the development of embedded iris biometric applications. ... level of convenience and security with unmatched biometric accuracy, ... identity aside from DNA. EyeLock,s platform uses video technology ...
Breaking Biology News(10 mins):