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


(Date:9/18/2017)... ... September 18, 2017 , ... Transportable biomass conversion facilities ... torrefied wood is the topic of a September 27 webinar hosted ... viability of transportable biomass conversion facilities for producing biochar, briquettes, and torrefied wood, ...
(Date:9/18/2017)... , Sept. 18, 2017   Montrium , a growing ... an exciting new partnership with a groundbreaking non-profit research organization, ... ... MDMA-assisted psychotherapy studies for PTSD ... has recently granted Breakthrough Therapy Designation to MDMA for the treatment ...
(Date:9/17/2017)... ... September 17, 2017 , ... GeneOne Life Science, Inc. announces ... Drug Safety (KMFDS) for an Investigational New Drug application for a Phase I/IIa ... (MERS-CoV). The study in Korea represents the second clinical trial for GLS-5300. ...
(Date:9/14/2017)... ... September 14, 2017 , ... ... Services, formally introduces its flexible scientist program (FSP)-- a flexible business approach ... The FSP, which combines SSCI’s extensive project-based analytical and solid-state chemistry services ...
Breaking Biology Technology:
(Date:4/3/2017)...  Data captured by IsoCode, IsoPlexis Corporation,s ... statistically significant association between the potency of ... objective response of cancer patients post-treatment. The ... cancer patients will respond to CAR-T cell ... to improve both pre-infusion potency testing and cell ...
(Date:3/30/2017)... 2017 Trends, opportunities and forecast in this ... technology (fingerprint, AFIS, iris recognition, facial recognition, hand geometry, ... end use industry (government and law enforcement, commercial and ... and others), and by region ( North America ... Asia Pacific , and the Rest of the ...
(Date:3/27/2017)... ROCKVILLE CENTRE, N.Y. , March 27, 2017 ... by Healthcare Information and Management Systems Society (HIMSS) ... Analytics Outpatient EMR Adoption Model sm . In ... top 12% of U.S. hospitals using an electronic ... recognized CHS for its high level of EMR ...
Breaking Biology News(10 mins):