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:2/8/2016)... Feb. 8, 2016  NanoViricides, Inc. (NYSE MKT: NNVC) (the "Company"), ... Eugene Seymour , MD, MPH, will present information about the company,s ... Hotel in New York City . ... the Windsor Room at 5:30PM EST. Registered attendees can request a ... York City . --> New York ...
(Date:2/8/2016)... , Feb. 8, 2016  Diplomat Pharmacy, Inc. (NYSE: DPLO) announced today that its new website ... pharmacy resource–user-centric, story-driven, knowledge-based and mobile-friendly. Visit the ... ... ... "The goal was to ...
(Date:2/6/2016)... ... 06, 2016 , ... The Center for Excellence in Education (CEE) will sponsor ... on Wednesday February 10, 2016. This Bite of Science session, hosted by the ... at 1500 Remount Road in Front Royal, VA from 5:00 p.m. to 8:00 p.m. ...
(Date:2/5/2016)... -- ATCC, the premier global biological materials resource and ... life science researchers that are working to address the ... CDC website . --> CDC website ... single-stranded RNA virus of the Flaviviridae family, genus Flavivirus, ... Viruses. Zika virus is transmitted to humans primarily through ...
Breaking Biology Technology:
(Date:2/3/2016)... 4, 2016 --> --> ... M (105.0), up 1,187% compared with fourth quarter of 2014. ... 517.6 M (loss: 30.0). Earnings per share increased to SEK ... 537.4 M (neg: 74.7). , --> ... to SEK 2,900.5 M (233.6), up 1,142% compared with 2014. ...
(Date:2/2/2016)... , Feb. 2, 2016  BioMEMS devices ... primarily focused on medical screening and diagnostic ... parameters. Wearable devices that facilitate and assure ... of movement are being bolstered through new ... biomedical signal acquisition coupled with wireless connectivity ...
(Date:2/2/2016)... , Feb. 2, 2016   Parabon ... from the U.S. Army Research Office and the ... range and sensitivity of the company,s Snapshot ... Accounting Mission and, more generally, defense-related DNA forensics. ... phenotyping capabilities (predicting appearance and ancestry from DNA ...
Breaking Biology News(10 mins):