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:10/10/2017)... (PRWEB) , ... October 10, 2017 , ... ... Science Center’s FirstHand program has won a US2020 STEM Mentoring Award. Representatives of ... award for Excellence in Volunteer Experience from US2020. , US2020’s mission is to ...
(Date:10/10/2017)... PA (PRWEB) , ... October 10, 2017 , ... ... year’s recipients of 13 prestigious awards honoring scientists who have ... presented in a scheduled symposium during Pittcon 2018, the world’s leading conference and ...
(Date:10/9/2017)... ... October 09, 2017 , ... ... journal, Epilepsia, Brain Sentinel’s SPEAC® System which uses the surface electromyography (sEMG) ... tonic-clonic seizures (GTCS) using surface electromyography (sEMG). The prospective multicenter phase III ...
(Date:10/6/2017)... ... October 06, 2017 , ... On ... and webinar on INSIGhT, the first-ever adaptive clinical trial for glioblastoma (GBM). The ... The event is free and open to the public, but registration is required. ...
Breaking Biology Technology:
(Date:4/5/2017)... 5, 2017  The Allen Institute for Cell Science ... a one-of-a-kind portal and dynamic digital window into the ... the first application of deep learning to create predictive ... lines and a growing suite of powerful tools. The ... and future publicly available resources created and shared by ...
(Date:4/3/2017)... 2017  Data captured by IsoCode, IsoPlexis ... a statistically significant association between the potency ... and objective response of cancer patients post-treatment. ... whether cancer patients will respond to CAR-T ... as to improve both pre-infusion potency testing and ...
(Date:3/29/2017)... the health IT company that operates the largest health ... today announced a Series B investment from BlueCross BlueShield ... investment and acquisition accelerates higi,s strategy to create the ... activities through the collection and workflow integration of ambient ... secures data today on behalf of over 36 million ...
Breaking Biology News(10 mins):