I am a student learning about genetic engineering and yeast transformation techniques. I don’t have research experience, but I want to understand what practical steps or methods can improve transformation efficiency in yeast. Any guidance or tips would be greatly appreciated.
Hello Shaik Afreen
Lithium acetate(LiAc)/single stranded DNA(ssDNA)/polyethylene glycol (PEG) is a standard method of yeast transformation. The transformation yield depends on the strain types and can be further increased using some minor changes.
- use fresh yeast colonies
- elute plasmid DNA/linear DNA/PCR cassette in sterile distilled water
- denature the ssDNA at 95 °C for 2 minutes followed by cool down before use
- wash pellet twice with 100 mM LiAc before adding transformation mixture to increase the competency
- prepare a transformation mixture (PEG, LiAc, ssDNA, DNA, water, if required) before adding to competent yeast cells and mix/vortex well
I am attaching a few transformation-related articles which may help you.
All the best!
See if you can find someone who does this protocol and ask if you can observe. Lots of lab protocols are written assuming the user knows things like "What sort of tube/container should I use?"; "How do I balance a centrifuge?"; "What do you mean by decant?"
Electroporation can definitely give better trasnformation efficiency than the Lithium acetate/PEG method
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