Navigation Links
Actinomyces viscosus

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.501 04/2002 Microorganism Actinomyces viscosus Cell type Bacteria, gram negative Molecules injected Plasmid DNA (in 1 mM Tris-HCl-0.1 mM EDTA (pH 8.0)) Growth medium Lactobacillus carrying medium (LCM) with 20 mM DL-threonine Washing solution Sterile, distilled water; sterile 10% glycerol Electroporation solution Sterile, 10% glycerol Outgrowth medium Complex medium (CAM) Cuvette 2 mm gap width Reference Yeung, M. K. and Kozelsky, C. S. 1994 Journal of Bacteriology 176. No. 13 4173-4176 Making electrocompetent cells:
  1. Grow a 500 ml cell culture in the early exponential phase until an O.D.600 of 0.2 is reached. Chill on ice for 1.5 h.
  2. 2. Harvest by centrifugation.
  3. Wash extensively first with a large volume of ice-cold sterile distilled water and then with sterile 10% glycerol.
  4. Resuspend in 10% glycerol to a final volume of 2 ml. Store in aliquots at 80 C.
Electroporation of cells:
  1. Thaw bacterial suspensions on ice. Add up to 5 l (100 ng) plasmid DNA to 100 l (5 x 108) of electrocompetent cells. Homogenize by gently mixing with pipette several times. Transfer mixture into 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. Dilute cells with 1 ml of CAM and incubate for 2 h at 37 C.
  5. Plate onto selective brain heart infusion agar plates; incubate at 37 C for up to 5 days.

Expected results:

Transformation efficiency up to 2.9 x 107 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Actinomyces viscosus
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/10/2017)... SANTA CRUZ, Calif. , Oct. 10, 2017 /PRNewswire/ ... SBIR grant from the NIH to develop RealSeq®-SC (Single ... preparation kit for profiling small RNAs (including microRNAs) from ... Cell Analysis Program highlights the need to accelerate development ... "New techniques for ...
(Date:10/9/2017)... ... ... The award-winning American Farmer television series will feature 3 Bar Biologics in an ... on RFD-TV. , With global population estimates nearing ten billion people by 2050, ... a growing nation. At the same time, many of our valuable resources are becoming ...
(Date:10/7/2017)... Phoenix, Arizona (PRWEB) , ... ... ... than 15 years’ experience providing advanced instruments and applications consulting for microscopy ... the in-house expertise in application consulting, Nanoscience Analytical offers a broad range ...
(Date:10/6/2017)... ... October 06, 2017 , ... On Tuesday, October 24th, ABC² ... the first-ever adaptive clinical trial for glioblastoma (GBM). The featured speaker will be ... and open to the public, but registration is required. , WHAT: ABC² ...
Breaking Biology Technology:
(Date:5/16/2017)... TEANECK, N.J. , May 16, 2017  Veratad ... leading provider of online age and identity verification solutions, ... the K(NO)W Identity Conference 2017, May 15 thru May ... Ronald Regan Building and International Trade Center. ... across the globe and in today,s quickly evolving digital ...
(Date:4/24/2017)... , April 24, 2017 ... and partner with  Identity Strategy Partners, LLP (IdSP) ... "With or without President Trump,s March 6, 2017 ... Terrorist Entry , refugee vetting can be instilled with ... resettlement. (Right now, all refugee applications are suspended ...
(Date:4/17/2017)... 17, 2017 NXT-ID, Inc. (NASDAQ: NXTD ... filing of its 2016 Annual Report on Form 10-K on Thursday ... ... available in the Investor Relations section of the Company,s website at ... website at http://www.sec.gov . 2016 Year Highlights: ...
Breaking Biology News(10 mins):