Navigation Links
Streptococcus salivarius

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.541 03/2002 Microorganism Streptococcus salivarius Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium HJGS (Hogg-Jago glucose broth with 0.4 sorbitol) Washing solution 5 mM potassium phosphate (pH 4.5), 0.4 M sorbitol, 10% glycerol Electroporation solution 5 mM potassium phosphate (pH 4.5), 0.4 M sorbitol, 10% glycerol Outgrowth medium HJGS (Hogg-Jago glucose broth with 0.4 sorbitol) Cuvette 1 mm gap width Reference Buckley, N. D. et al 1999 Applied and Environmental Microbiology 65, No. 9 3800-3804 Making electrocompetent cells:

1. Cultivate cells by adding 1% overnight preculture to fresh medium. Grow cells at 37 C without agitation to a cell
density of O.D.660 of 0.5. Add glycine to a final concentration of 10% and incubate again for 1 h at 37 C. 2. Harvest by centrifugation. 3. Wash twice in ice-cold washing solutio n. 4. Resuspend in electroporation solution to a 50-fold concentration and freeze in an ethanol-dry ice bath. Store at -80 C.

Electroporation of cells:

  1. Thaw cells on ice. Add 1 g plasmid DNA to 100 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) 1,600 V Time constant (T) 5 ms
  4. Add 1 ml ice-cold outgrowth medium and incubate for 3 h at 37 C.
  5. Plate aliquots onto selective agar plates; incubate 2-3 days at 37 C.
Expected Results: Transformation efficiency up to 1 x 105 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Streptococcus salivarius
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/11/2017)... ... October 11, 2017 , ... ... has granted orphan drug designation to SBT-100, its novel anti-STAT3 (Signal Transducer and ... of osteosarcoma. SBT-100 is able to cross the cell membrane and bind intracellular ...
(Date:10/10/2017)... 2017 International research firm Parks Associates announced today ... the TMA 2017 Annual Meeting , October 11 in ... home security market and how smart safety and security products impact the ... Parks Associates: Smart Home Devices: ... "The residential security market has experienced ...
(Date:10/9/2017)... Jupiter, FL (PRWEB) , ... October 09, 2017 , ... ... episode, scheduled to broadcast first quarter 2018. American Farmer airs Tuesdays at 8:30aET on ... Agriculture industry is faced with the challenge of how to continue to feed a ...
(Date:10/7/2017)... ... October 06, 2017 , ... ... its ProxiMeta™ Hi-C metagenome deconvolution product, featuring the first commercially available Hi-C ... software to perform Hi-C metagenome deconvolution using their own facilities, supplementing the ...
Breaking Biology Technology:
(Date:3/30/2017)... -- Trends, opportunities and forecast in this market to ... AFIS, iris recognition, facial recognition, hand geometry, vein recognition, ... industry (government and law enforcement, commercial and retail, health ... and by region ( North America , ... , and the Rest of the World) ...
(Date:3/27/2017)... , March 27, 2017  Catholic Health ... and Management Systems Society (HIMSS) Analytics for achieving ... Adoption Model sm . In addition, CHS previously ... U.S. hospitals using an electronic medical record (EMR). ... its high level of EMR usage in an ...
(Date:3/23/2017)... The report "Gesture Recognition and Touchless Sensing Market by Technology (Touch-based ... to 2022", published by MarketsandMarkets, the market is expected to be worth USD ... 2022. Continue Reading ... ... ...
Breaking Biology News(10 mins):