PEG mediated plasmid DNA transformation into protoplasts?

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Jong-hwan Shin

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.

Simran Bhullar

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,

Challagandla Anil

Thank you@ Shin and Bhullar.

These protocols can work with filamentous fungi (Colletotrichum truncatum)