Does anyone have a detailed protocol for PEG mediated plasmid DNA transformation? Which MW should PEG have for a plasmid of 8.6kb?
Thank you in advance.
This is a protocol for fungal transformation.
Disolve 8g of PEG 3350 in 2 ml of water. ( final volume 10 ml )
Prepare 2X STC.
Add 3~5 ug of plasmid DNA to 200 ul of protoplast.
Wait for 10minutes.
Mix 500 ul of PEG with 500 ul of 2X STC.
Add 1 ml of PTC ( PEG + STC ).
Wait for 20minutes.
Incubate in a shaker for 6 hours at appropriate RPM and regenerate
with appropriate media and selected antibiotics.
We have done this and you can see the protocol in the paper: http://www.plantphysiol.org/content/132/2/988.full
We used PEG 1450.
The paper is available on my profile as well.
Best,
S
Thank you@ Shin and Bhullar.
These protocols can work with filamentous fungi (Colletotrichum truncatum)
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