Navigation Links
Mycobacterium avium

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.523 03/2002 Microorganism Mycobacterium avium 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 intracellulare
2. Mycobacterium smegmatis
3. Mycobacterium tuberculosis
4. Mycobacterium avium
5. Mycobacterium intracellulare
6. Mycobacterium smegmatis
7. Mycobacterium tuberculosis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/10/2017)... ... October 10, 2017 , ... For the second ... a US2020 STEM Mentoring Award. Representatives of the FirstHand program travelled to Washington, ... from US2020. , US2020’s mission is to change the trajectory of STEM education ...
(Date:10/10/2017)... SomaGenics announced the receipt of a Phase ... (Single Cell), expected to be the first commercially available ... from single cells using NGS methods. The NIH,s recent ... development of approaches to analyze the heterogeneity of cell ... for measuring levels of mRNAs in individual cells have ...
(Date:10/9/2017)... SAN DIEGO , Oct. 9, 2017 /PRNewswire/ ... a biological mechanism by which its ProCell stem ... of critical limb ischemia.  The Company, demonstrated that ... the amount of limbs saved as compared to ... of the molecule HGF resulted in reduction of ...
(Date:10/9/2017)... ... October 09, 2017 , ... The Giving Tree Wellness ... targeting the needs of consumers who are incorporating medical marijuana into their wellness ... Arizona. , As operators of two successful Valley dispensaries, The Giving Tree’s two ...
Breaking Biology Technology:
(Date:4/17/2017)... Florida , April 17, 2017 NXT-ID, ... technology company, announces the filing of its 2016 Annual Report on ... and Exchange Commission. ... on Form 10-K is available in the Investor Relations section of ... as on the SEC,s website at http://www.sec.gov . ...
(Date:4/13/2017)... 2017 According to a new market research report ... Identity Administration, and Authorization), Service, Authentication Type, Deployment Mode, Vertical, and Region ... expected to grow from USD 14.30 Billion in 2017 to USD 31.75 ... ... MarketsandMarkets Logo ...
(Date:4/11/2017)... DUBLIN , Apr. 11, 2017 Research ... Tracking Market 2017-2021" report to their offering. ... The global eye tracking market to grow at ... The report, Global Eye Tracking Market 2017-2021, has been prepared based ... report covers the market landscape and its growth prospects over the ...
Breaking Biology News(10 mins):