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 study ... in frozen and fresh in vitro fertilization (IVF) transfer cycles. The ... IVF success. , After comparing the results from the fresh and frozen transfer ...
(Date:10/10/2017)... Poway, California (PRWEB) , ... October 10, 2017 , ... ... afternoon speaking at his local San Diego Rotary Club. The event ... San Diego, CA and had 300+ attendees. Dr. Harman, DVM, MPVM was joined ...
(Date:10/9/2017)... DIEGO , Oct. 9, 2017  BioTech ... biological mechanism by which its ProCell stem cell ... critical limb ischemia.  The Company, demonstrated that treatment ... amount of limbs saved as compared to standard ... the molecule HGF resulted in reduction of therapeutic ...
(Date:10/7/2017)... (PRWEB) , ... October 06, ... ... years’ experience providing advanced instruments and applications consulting for microscopy and surface ... expertise in application consulting, Nanoscience Analytical offers a broad range of contract ...
Breaking Biology Technology:
(Date:8/23/2017)... ARMONK, N.Y. , Aug. 23, 2017  The general public,s help ... the human microbiome—the bacteria that live in and on the human body ... ... bacteria in the human microbiome, starting with the gut. The project's goal ... in disease. Photo credit: IBM ...
(Date:7/20/2017)... , July 20, 2017 Delta (NYSE: DAL ) ... any Delta aircraft at Reagan Washington National Airport (DCA). ... Delta launches biometrics to board ... Delta,s biometric boarding ... Club is now integrated into the boarding process to allow eligible ...
(Date:6/23/2017)... and ITHACA, N.Y. , June 23, ... University, a leader in dairy research, today announced a ... to help reduce the chances that the global milk ... of this dairy project, Cornell University has become the ... the Food Supply Chain, a food safety initiative that ...
Breaking Biology News(10 mins):