Navigation Links
Mycobacterium smegmatis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.522 03/2002 Microorganism Mycobacterium smegmatis LR222 Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium Middlebrook 7H9 medium with 0.1% Tween and Dubos oleic albumin complex
enrichment Washing solution 10% glycerol Electroporation solution 10% glycerol Outgrowth medium Middlebrook 7H9 liquid medium with 0.1% Tween and Dubos oleic albumin complex
enrichment, selective Middlebrook 7H10 agar plates Cuvette 1 mm gap width Reference Beggs, M. L.. et al 1995 Journal of Bacteriology 177, No. 17 4836-4840 Making electrocompetent cells:

1. Grow cells in sidearm flask or tissue culture plates with gentle agitation until a Klett unit reading of 100 to 200 was
obtained. 2. Harvest cells and wash three times with cold sterile 10% glycerol. 3. Resuspend 1/100 volume of 10% glycerol.

Electroporation of cells:

  1. Add up to 1 g plasmid DNA to 60 l of electrocompetent cells. Homogenize by gently mixing with pipette several
    times. Incubate on ice for 10 min. Transfer mixture into a prechilled cuvette.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 1,200 V Time constant (T) 5 ms
  4. Wash cells out of the cuvette with 1 ml of outgrowth medium and incubate for 2 h at 37 C.
  5. Plate on selective agar.
Expected Results: Transformation efficiency up to 105 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Mycobacterium avium
2. Mycobacterium intracellulare
3. Mycobacterium tuberculosis
4. Mycobacterium avium
5. Mycobacterium intracellulare
6. Mycobacterium smegmatis
7. Mycobacterium tuberculosis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:5/26/2016)... May 26, 2016 Q BioMed Inc. ... will be a featured presenter at the 5th Annual Marcum ... New York City at the Grand Hyatt Hotel. ... Q BioMed Inc. CEO, is scheduled to begin at 11a.m ... company,s business strategy, recent developments and outline milestones for the ...
(Date:5/25/2016)... ... May 25, 2016 , ... Lajollacooks4u has become a rising hotspot for ... one of its top attractions. Fortune 500 companies, such as Illumina, Hewlett-Packard, Qualcomm ... and intimate team-building experience. , Each event kicks off with an olive oil and ...
(Date:5/25/2016)... ... ... at the University of Athens say they have evidence that the variety of different ... lead to one good one. Surviving Mesothelioma has just posted an article on the ... 98 mesothelioma patients who got a second kind of drug therapy after ...
(Date:5/24/2016)... 24, 2016   MedyMatch Technology Ltd ., the data ... real-time decision support tools in the emergency room, announced today ... Israeli Advanced Technology Industries (IATI) BioMed Conference. ... 15th National Life Sciences and Technology Week, and is ... Hotel in Tel Aviv, Israel . ...
Breaking Biology Technology:
(Date:3/10/2016)... March 10, 2016   Unisys Corporation (NYSE: ... Protection (CBP) is testing its biometric identity solution at ... to help identify certain non-U.S. citizens leaving the ... test, designed to help determine the efficiency and accuracy of ... February and will run until May 2016. --> ...
(Date:3/9/2016)... BEACH GARDENS, Fla. , March 9, 2016 ... identity management authentication and enrollment solutions, today announced ... DigitalPersona ® Altus multi-factor authentication ... IT and InfoSec managers to step-up security where ... Washington, DC . ...
(Date:3/3/2016)... , March 3, 2016  2016FLEX, organized by ... week highlighting advancements in flexible, hybrid and printed ... setting attendance - have gathered for short courses, ... field of electronics. The Flex Conference celebrates its ... for companies, R&D organizations, and universities contributing to ...
Breaking Biology News(10 mins):