Navigation Links
Nocardia corallina

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.526 01/2002 Microorganism Nocardia corallina Cell type Bacteria, gram positive Molecules injected Plasmid DNA (in TE buffer) Growth medium N2 medium (20 g/l glucose, 2 g/l yeast extract, 4 g/l beef extract, 6 g/l tryptone, 2 g/l NaCl, 10 g/l glycine) Washing solution Distilled water Electroporation solution 0.3 M sucrose, 15% glycerol Outgrowth medium N2 medium (without antibiotics) Cuvette 2 mm gap width Reference Yao W., et al 1994 Current Microbiology 29 223-227 Making electrocompetent cells:

1. Inoculate a fresh overnight culture of bacteria into N2 medium. Grow cells at 28 C to an O.D.600 of 0.7. Harvest by centrifugation.. 2. Wash twice with _ volume of distilled water. 3. Resuspend in a final volume of 1/20 of the original volume of electroporation solution.

Electroporation of cel ls:

  1. Add 2 l (0.1 g) plasmid DNA to 50 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. Add 1 ml N2 medium and incubate for 2 hours at 30 C.
  5. Plate cells on selective YEME medium with a 3 ml YEME soft agar (0.4%) overlayment.
Expected Results: Transformation efficiency up to 2.8 x 103 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Nocardia asteroides
2. Nocardia corallina
3. Nocardia asteroides
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/11/2017)... ... October 11, 2017 , ... A new ... rates in frozen and fresh in vitro fertilization (IVF) transfer cycles. ... to IVF success. , After comparing the results from the fresh and frozen ...
(Date:10/10/2017)... Philadelphia, PA (PRWEB) , ... October 10, 2017 ... ... University City Science Center’s FirstHand program has won a US2020 STEM Mentoring Award. ... accept the award for Excellence in Volunteer Experience from US2020. , US2020’s mission ...
(Date:10/10/2017)... ... October 10, 2017 , ... ... process optimization firm for the life sciences and healthcare industries, announces a presentation ... San Francisco. , The presentation, “Automating GxP Validation for Agile Cloud Platforms,” will ...
(Date:10/9/2017)... , ... October 09, 2017 , ... At its national ... Christopher Stubbs, a professor in Harvard University’s Departments of Physics and Astronomy, has been ... a member of the winning team for the 2015 Breakthrough Prize in Fundamental physics ...
Breaking Biology Technology:
(Date:4/5/2017)... 2017  The Allen Institute for Cell Science today ... one-of-a-kind portal and dynamic digital window into the human ... first application of deep learning to create predictive models ... and a growing suite of powerful tools. The Allen ... future publicly available resources created and shared by the ...
(Date:3/30/2017)... , March 30, 2017  On April 6-7, 2017, ... the Genome hackathon at Microsoft,s headquarters in ... competition will focus on developing health and wellness apps ... Hack the Genome is the first hackathon ... The world,s largest companies in the genomics, tech and ...
(Date:3/27/2017)... 2017  Catholic Health Services (CHS) has been ... (HIMSS) Analytics for achieving Stage 6 on the ... In addition, CHS previously earned a place in ... electronic medical record (EMR). "HIMSS Analytics ... EMR usage in an outpatient setting.  This recognition ...
Breaking Biology News(10 mins):