Difference between revisions of "Protoplast Transformation"

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(Created page with "#Follow the instruction in Protoplasts Formation, Regeneration and Transformation #Add 3.85 ml P buffer to 1 g PEG sterilised. #Add 200 µl PEG-P buffer mixture and a DNA...")
 
(Materials)
 
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==Materials==
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*[[P buffer]]
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*1 g polyethylene glycol 1000 (PEG)
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*[[R5]] or [[R2YE]] plate
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==Instructions==
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#Follow the instruction in [[Protoplasts Formation, Regeneration and Transformation]]
 
#Follow the instruction in [[Protoplasts Formation, Regeneration and Transformation]]
 
#Add 3.85 ml P buffer to 1 g PEG sterilised.
 
#Add 3.85 ml P buffer to 1 g PEG sterilised.

Latest revision as of 22:58, 24 March 2020

Materials

Instructions

  1. Follow the instruction in Protoplasts Formation, Regeneration and Transformation
  2. Add 3.85 ml P buffer to 1 g PEG sterilised.
  3. Add 200 µl PEG-P buffer mixture and a DNA solution to 50 µl of the protoplast suspension.
  4. Plate it onto R5 or R2YE.
  5. Incubate the plate at 298-30ºC overnight.
  6. Overlay 1 ml sterilised water containing nalidixic acid and an appropriate antibiotic.