Navigation Links
Kluyveromyces lactis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.516 12/2001 Microorganism Kluyveromyces lactis AWJ137 Cell type Yeast Molecules injected DNA Growth medium Not given Washing solution YPD, 10 mM DTT, 40 mM HEPES (pH 8.0) Electroporation solution 2.5 mM Tris/HCl, 1 mM MgCl2, (pH 8.0) Outgrowth medium YPD with 1 M sorbitol Cuvette 4 mm gap width Reference Prof. Friedhelm Meinhardt Universitt Mnster Corrensstr. 3 D-48149 Mnster
e-mail: meinhar@uni-muenster.de Making electrocompetent cells:

1. Grow a 50 ml cell culture of Kluyveromyces lactis to a density of 5 x 107 cells/ml. 2. Harvest by centrifugation (5 min., at 4,000 x g) 3. Resuspend cells in 5 ml washing solution and incubate for 5 min. 4. Wash twice with electroporation solution and resuspend in 0.25 ml of the same solution. Incubate for 1 hour on ice.

Electroporation of cells:

  1. Add 5 l DNA (1 g/l) 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) 2,500 V Time constant (T) 5 ms
  4. Add 1 ml YPD medium with 1 M sorbitol, incubate for 1 hour at 30 C.
  5. Centrifuge cells 30 sec., resuspend in 0.4 ml sterile water, plate in aliquots on selective plates.
Expected results: Transformation efficiency up to 200 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Kluyveromyces lactis
2. Lactococcus lactis
3. Lactococcus lactis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/11/2017)... ... , ... Disappearing forests and increased emissions are the main causes of the ... Especially those living in larger cities are affected by air pollution related diseases. , ... pollution-affected countries globally - decided to take action. , “I knew I had to ...
(Date:10/11/2017)... (PRWEB) , ... October 11, 2017 , ... ... Administration (FDA) has granted orphan drug designation to SBT-100, its novel anti-STAT3 (Signal ... the treatment of osteosarcoma. SBT-100 is able to cross the cell membrane and ...
(Date:10/10/2017)... ... October 10, 2017 , ... ... antibody-drug conjugate (ADC) therapeutics, today confirmed licensing rights that give it exclusive ... a technology developed in collaboration with Children’s Hospital Los Angeles (CHLA). ...
(Date:10/10/2017)... California (PRWEB) , ... October 10, 2017 , ... Dr. ... speaking at his local San Diego Rotary Club. The event entitled ... Diego, CA and had 300+ attendees. Dr. Harman, DVM, MPVM was joined by ...
Breaking Biology Technology:
(Date:4/3/2017)... April 3, 2017  Data captured by ... platform, detected a statistically significant association between ... to treatment and objective response of cancer ... to predict whether cancer patients will respond ... as well as to improve both pre-infusion potency ...
(Date:3/30/2017)... -- The research team of The Hong Kong Polytechnic ... by adopting ground breaking 3D fingerprint minutiae recovery and matching technology, ... accuracy for use in identification, crime investigation, immigration control, security of ... ... A research team led by Dr ...
(Date:3/29/2017)... -- higi, the health IT company that operates the largest ... , today announced a Series B investment from BlueCross ... new investment and acquisition accelerates higi,s strategy to create ... health activities through the collection and workflow integration of ... and secures data today on behalf of over 36 ...
Breaking Biology News(10 mins):