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:10/11/2017)... ... 11, 2017 , ... Personal eye wash is a basic first aid supply for any work ... which eye do you rinse first if a dangerous substance enters both eyes? It’s one ... with its unique dual eye piece. , “Whether its dirt and debris, or an ...
(Date:10/11/2017)... ... 11, 2017 , ... Disappearing forests and increased emissions are the main causes ... each year. Especially those living in larger cities are affected by air pollution related ... the most pollution-affected countries globally - decided to take action. , “I knew I ...
(Date:10/10/2017)... ... October 10, 2017 , ... For the second time in ... STEM Mentoring Award. Representatives of the FirstHand program travelled to Washington, D.C. Tuesday, ... , US2020’s mission is to change the trajectory of STEM education in America ...
(Date:10/10/2017)... ... 10, 2017 , ... The Pittcon Program Committee is pleased ... scientists who have made outstanding contributions to analytical chemistry and applied spectroscopy. Each ... leading conference and exposition for laboratory science, which will be held February 26-March ...
Breaking Biology Technology:
(Date:4/11/2017)... 2017 Research and Markets has announced the ... to their offering. ... eye tracking market to grow at a CAGR of 30.37% during ... Market 2017-2021, has been prepared based on an in-depth market analysis ... and its growth prospects over the coming years. The report also ...
(Date:4/5/2017)... , April 5, 2017  The Allen Institute ... Allen Cell Explorer: a one-of-a-kind portal and dynamic digital ... 3D imaging data, the first application of deep learning ... human stem cell lines and a growing suite of ... platform for these and future publicly available resources created ...
(Date:4/3/2017)... WASHINGTON , April 3, 2017 /PRNewswire-USNewswire/ ... single-cell precision engineering platform, detected a statistically ... cell product prior to treatment and objective ... highlight the potential to predict whether cancer ... prior to treatment, as well as to ...
Breaking Biology News(10 mins):