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:3/29/2017)... Germany , March 29, 2017 QIAGEN ... STANDARD: QIA) today announced the U.S. launch of its ipsogen ... cleared by the U.S. Food and Drug Administration as a qualitative in vitro ... extracted from EDTA whole blood.* ... ipsogen JAK2 assay is processed on QIAGEN,s Rotor-Gene ® Q MDx ...
(Date:3/29/2017)... TIKVAH, Israel , March ... (NASDAQ: BCLI), a leading developer of adult ... financial results for the year ended December ... highly successful and pivotal year for Brainstorm, with a ... made on clinical, regulatory and operational fronts," said ...
(Date:3/29/2017)... , March 29, 2017  Halozyme Therapeutics, Inc. ... and drug-delivery therapies, today announced that an Oncologic ... Drug Administration voted 11 to 0 that the ... injection was favorable for patients in the proposed ... and chronic lymphocytic leukemia. The FDA action date ...
(Date:3/29/2017)... ... March 29, 2017 , ... Bactana Animal Health, a ... the world’s food supply through enhancement of the gut microbiota, today announced the closing ... Connecticut Innovations , New York-based Sustainable Income Capital Management, LLC and a number of ...
Breaking Biology Technology:
(Date:2/14/2017)... Feb. 14, 2017  Wake Forest Baptist Medical Center ... new chief executive officer (CEO). Freischlag joins the medical ... D. McConnell , M.D., who last year announced that ... Medical Center, after leading it since 2008.   ... of Wake Forest Baptist,s academic health system, which includes ...
(Date:2/13/2017)... WASHINGTON , Feb. 13, 2017 Former ... U.S. Senate Judiciary Committee, Janice Kephart of ... regarding President Donald Trump,s "Executive Order: Protecting ... States" (Jan. 27, 2017):  "As President Trump,s ... 9th Circuit has now essentially banned the travel ban, ...
(Date:2/9/2017)... LONDON , Feb. 9, 2017 The ... in-depth analysis of the biomass boiler market globally in ... sales of biomass boilers. The market for biomass boilers ... product type, end-user, application, and country/region. The market based ... agriculture & forest residues, biogas & energy crops, urban ...
Breaking Biology News(10 mins):