Navigation Links
Mycobacterium tuberculosis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.524 03/2002 Microorganism Mycobacterium tuberculosis H37Rv Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium Middlebrook 7H9 medium with 0.2% glycerol, 0.25% Tween 80, albumin-dextrose complex (ADC) Washing solution 10% glycerol Electroporation solution 10% glycerol Outgrowth medium Middlebrook 7H9 medium with ADC, selective 7H10 agar plates with ADC Cuvette 2 mm gap width Reference Armitige, L. Y. et al 2000 Infection and Immunity 68, No. 2 767-778 Making electrocompetent cells:

1. Grow cells with gentle shaking to an OD600 of 0.6-1.0. 2. Harvest cells and wash three times in 1/50 volume of cold 10% glycerol. 3. Resuspend in 10% glycerol at a concentration of approx. 1011 cells/ml and store at 70 C until needed.

Electroporation of cells:

  1. Add 2-4 g plasmid DNA to 100 l (1010) 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) 1,200 V Time constant (T) 5 ms
  4. Immediately add 1 ml outgrowth medium and incubate at 37 C for 2.5 h with agitation.
  5. Plate on selective agar plates and incubate at 37 C for 3 weeks under 9.3% CO2.
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 smegmatis
4. Mycobacterium avium
5. Mycobacterium intracellulare
6. Mycobacterium smegmatis
7. Mycobacterium tuberculosis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:5/24/2016)... --  MedyMatch Technology Ltd ., the data analytics healthcare company ... tools in the emergency room, announced today that it has ... Industries (IATI) BioMed Conference. The Conference is ... Life Sciences and Technology Week, and is being held May ... Tel Aviv, Israel . Gene Saragnese , ...
(Date:5/23/2016)... ... May 23, 2016 , ... The need for blood donations in South Texas and ... the South Texas Blood & Tissue Center, blood donations are on the decline. In fact, ... they are down 21 percent in South Texas in the last four years alone. , ...
(Date:5/23/2016)... -- Oxitec CEO Hadyn Parry will ... ET before the United States House Committee on Science, Space ... in controlling the spread of the Aedes aegypti ...      (Logo: http://photos.prnewswire.com/prnh/20150630/227348 ) Oxitec has ... Trials in Brazil , Panama ...
(Date:5/23/2016)... Alto, CA (PRWEB) , ... May 23, 2016 ... ... and public interest organization focused on molecular nanotechnology, announced the winners for the ... of pioneer physicist Richard Feynman, are given in two categories, one for experiment ...
Breaking Biology Technology:
(Date:5/12/2016)... , May 12, 2016 WearablesResearch.com ... just published the overview results from the Q1 wave ... the recent wave was consumers, receptivity to a program ... data with a health insurance company. "We ... to share," says Michael LaColla , CEO of ...
(Date:4/28/2016)... , April 28, 2016 Infosys ... (NYSE: INFY ), and Samsung SDS, a global ... that will provide end customers with a more secure, fast ...      (Logo: http://photos.prnewswire.com/prnh/20130122/589162 ) , ... but it also plays a fundamental part in enabling and ...
(Date:4/26/2016)... , April 27, 2016 ... "Global Multi-modal Biometrics Market 2016-2020"  report to their ... , The analysts forecast the global ... of 15.49% during the period 2016-2020.  ... of sectors such as the healthcare, BFSI, transportation, ...
Breaking Biology News(10 mins):