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Making Yeast Competent Cells and Yeast Cell Transformation
Published: Jul 20, 2011 DOI: 10.21769/BioProtoc.96 Views: 41068
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Abstract
This is a quite simple but reliable protocol to make very high transformation efficiency yeast competent cells. By express your gene of interest, protein function can be studied in yeast cells.
Materials and Reagents
- Bacto-Yeast extract (Thermo Fisher Scientific)
- Bacto-peptone (Thermo Fisher Scientific)
- Glucose (dextrose) (Thermo Fisher Scientific)
- Bacto-agar (Thermo Fisher Scientific)
- Deionized H2O
- Glycerol (Sigma-Aldrich)
- Dimethyl sulfoxide (DMSO) (Sigma-Aldrich)
- PEG 3350 (Sigma-Aldrich)
- Lithium acetate dihydrate (LiAc) (Sigma-Aldrich)
- Salmon sperm DNA (Life Technologies, Invitrogen™)
- Yeast nitrogen base (YNB) (Sigma-Aldrich)
- 2-(N-morpholino) ethanesulfonic acid (MES)
- ADE (Sigma-Aldrich)
- Agar
- YPAD plate & liquid medium (see Recipes)
- Transformation solution (see Recipes)
- YNB + MA plate (see Recipes)
Equipment
- Water bath (VWR International)
- Centrifuges (Eppendorf)
- 30 °C shaker and incubator (VWR International)
- Standard petri dishes (VWR International)
- 1.5 ml centrifuge tubes (Eppendorf)
Procedure
Category
Cell Biology > Cell isolation and culture > Transformation
Microbiology > Microbial genetics > Transformation
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