Navigation Links
Mycobacterium intracellulare

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.521 02/2002 Microorganism Mycobacterium intracellulare 1403 Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium Middlebrook 7H9 liquid medium with oleic acid dextrose complex (OADC) Washing solution 10% glycerol Electroporation solution 10% glycerol Outgrowth medium Middlebrook 7H9 liquid medium with oleic acid dextrose complex Cuvette 2 mm gap width Reference Marklund, B.-I. et al 1995 Journal of Bacteriology 177, No. 21 6100-6105 Making electrocompetent cells:

1. Grow a 50-100 ml cell culture at 37 C statically to an O.D.600 of 0.1 to 0.3. 2. Harvest by centrifugation. 3. Wash twice in cold 10% glycerol. The first time in 30 ml the second time in 1 ml. 4. Resuspend 0.2-0.5 ml of 10% glycerol.

Electroporation of cells:< /b>

  1. Add 1 g plasmid DNA to 100 l of electrocompetent cells. Homogenize by gently mixing with pipette several times. 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) 2,500 V Time constant (T) 5 ms
  4. Transfer cells to 1 ml of 7H9 medium and grow statically for 16-20 h.
  5. To disperse cells, bath sonicate cultures twice for 30 s each time. Dilute suspension in 0.9% NaCl supplemented with 0.1% Tween 80.
  6. Plate on selective agar.
Expected Results: Transformation efficiency up to 106 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

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


(Date:1/18/2017)... Jan. 18, 2017 Shareholder rights law firm ... whether the board members of CoLucid Pharmaceuticals, Inc. (NASDAQ: ... connection with the proposed sale of the Company to ... that develops small molecules for the acute treatment of ... it had signed a definitive merger agreement with Eli ...
(Date:1/18/2017)...  Market Research Future published a half-cooked research report on Global ... at a CAGR of 12% during the period 2016 to 2022. ... ... cell division without any control. These abnormal cells have the ability ... cells can spread to other parts of the body through the ...
(Date:1/17/2017)...  An international team of researchers from UC ... Hospital Albrechtsen Research Centre/University of Manitoba have identified ... need affecting nearly one in 15 Americans. Published ... results identify small molecule drugs with neuroprotective and ... in animal models of metabolic, chemical and infection ...
(Date:1/17/2017)... , Jan. 17, 2017 Noom ... is the first to offer fully Spanish behavior ... Noom,s Spanish diabetes prevention and healthy ... and accessibility of lifestyle interventions among Hispanic Americans ... conditions. Noom,s robust food database, program-specific curriculum content ...
Breaking Biology Technology:
(Date:12/7/2016)... BioCatch , the global leader in behavioral ... which grew to over 40 granted and pending patents. ... , , The Company,s ... " System, Device, and Method Estimating Force Applied to a Touch ... forego costly hardware components needed to estimate the force and pressure applied ...
(Date:12/6/2016)... , Dec. 6, 2016  Zimmer Biomet Holdings, Inc. (NYSE ... priced an offering of €500.0 million principal amount of its ... amount of its 2.425% senior unsecured notes due 2026. ... occur on December 13, 2016, subject to the satisfaction of customary ... basis. The Company intends to ...
(Date:12/5/2016)...  The Office of Justice Programs, National Institute ... Enhance or Replace Medico Legal Autopsies?" on NIJ.gov.  ... replacing forensic autopsies with postmortem X-ray computed tomography, ... response to recommendations made by The National Academy ... as a potential component of medicolegal death investigations. ...
Breaking Biology News(10 mins):