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


(Date:1/13/2017)... Research and Markets has announced the addition of the "Global ... ... CAGR of 16.83% during the period 2017-2021. The report ... market for 2017-2021. To calculate the market size, the report considers the ... also includes a a discussion of the key vendors operating in this ...
(Date:1/12/2017)... (PRWEB) , ... January 12, ... ... USING VTI’S INTERFUSE® IS A SUCCESS , VTI, Vertebral Technologies, Inc., announces ... the InterFuse® laterally expandable device. Since September 2016, VTI (Vertebral Technologies, Inc.) ...
(Date:1/12/2017)... ... January 12, 2017 , ... ... treatments, 26-year-old Lisa Rosendahl’s doctors gave her only a few months to live. ... drug combination that has stabilized Rosendahl’s disease and increased both the quantity and ...
(Date:1/12/2017)... -- The report "Direct-Fed Microbials Market by Type (Lactic Acid Bacteria and Bacillus), ... - Global Forecast to 2022", published by MarketsandMarkets, the global market is estimated ... USD 1,399.6 Million by 2022, at a CAGR of 6.96% from 2016. ... ... MarketsandMarkets Logo ...
Breaking Biology Technology:
(Date:12/15/2016)... 2016   WaferGen Bio-systems, Inc. (NASDAQ: ... announced today that on December 13, 2016, it received ... Nasdaq Stock Market LLC which acknowledged that, as of ... common stock had been at $1.00 or greater for ... with Listing Rule 5550(a)(2) of the Nasdaq Stock Market. ...
(Date:12/15/2016)... "Increase in mobile transactions is driving the ... market is expected to grow from USD 4.03 billion ... a CAGR of 29.3% between 2016 and 2022. The ... demand for smart devices, government initiatives, and increasing penetration ... is expected to grow at a high rate during ...
(Date:12/7/2016)... According to a new market research report "Emotion Detection and Recognition ... Service, Application Area, End User, And Region - Global Forecast to 2021", published ... in 2016 to USD 36.07 Billion by 2021, at a Compound Annual Growth ... ... MarketsandMarkets Logo ...
Breaking Biology News(10 mins):