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Cat. No.: PTSC-50 (for 50T)

Cat. No.: PTSC-250 (for 250T)

Description

Competent Cell Preparation and Transformation Kit for Saccharomyces Cerevisiae is a fast, convenient, and efficient kit for preparing highly active yeast competent cells and performing plasmid transformation. The kit provides almost all the reagents needed for competent cell preparation and transformation, eliminating the need for dilution and preparation. The process is simple and quick, with competent cells ready within 30 minutes after yeast culture completion. It can be used for yeast hybridization experiments and yeast library construction experiments.

During yeast transformation, yeast cells are cultured using the buffer solution provided in the kit to bring them to a competent state. The competent cells are then mixed with the plasmid DNA and Carrier DNA before incubating with the transformation solution. Carrier DNA, a short single-stranded linear DNA, facilitates the uptake of exogenous plasmid DNA by yeast cells and protects the plasmid from DNA enzyme degradation.

Saccharomyces cerevisiae, commonly known as brewer's yeast, is a single-celled eukaryotic microorganism belonging to the genus Saccharomyces. It is extensively studied and used as a model organism in genetics. Its genome is approximately 1.2×107 base pairs long, containing 16 chromosomes with around 6000 open reading frames (ORFs). Only 4% of yeast genes have introns, making its genetic background relatively simple. Brewer's yeast exhibits growth characteristics similar to prokaryotes, making it easy to culture and genetically manipulate. It is considered a model eukaryotic organism and often referred to as the "E. coli of eukaryotes". Brewer's yeast can exist in haploid form, facilitating genotype-phenotype analysis and efficient homologous recombination for genome editing and high-throughput genetic analysis. Its expression system processes exogenous genes to some extent post-translationally, enabling proper folding and glycosylation modification of harvested foreign proteins, which enhances protein activity and stability. Additionally, exogenous genes can be secreted when expressed in brewer's yeast, facilitating purification and preventing intracellular accumulation of the expressed product.

During brewer's yeast transformation, appropriate plasmids can be selected based on the nutritional deficiencies of the strains. Generally, if a mutant strain lacks specific components in the culture medium (amino acids, purines, or pyrimidines), it cannot grow. Using plasmids that complement the mutant genes allows the transformants to grow on media lacking specific components. Typically, transforming 1µg of plasmid can yield >103 transformants, although transformation efficiency may vary between different strains of brewer's yeast.

Components

• Buffer A
• Buffer B
• Buffer C
• Carrier DNA
• DMSO

Storage

Store at -20ºC, effective for at least one year. Except for Carrier DNA, store at 4ºC, effective for three months.

Precaution

• When using Carrier DNA, the required amount can be heated at 100ºC for 15 minutes according to the transformation quantity. Then, immediately place it in an ice bath for 15 minutes before use. Any excess after use can be stored at -20ºC, and when needed again, thaw the Carrier DNA on ice and boil it again to denature. If the amount used each time is small, it can be divided appropriately to avoid repeated freeze-thaw cycles. Heating treatment can shorten and denature the Carrier DNA into single strands, and immediate cooling in an ice bath maintains it in a single-stranded state. It is not recommended to heat all the Carrier DNA at 100ºC for 15 minutes at once, as multiple treatments may make the Carrier DNA excessively short.
• The volume of samples for transformation should not be too large, as larger volumes can lead to decreased transformation efficiency.
• Competent cells are very sensitive to temperature changes and should avoid temperature changes not specified in the instructions.
• Competent cells are also sensitive to mechanical force. When adding samples for transformation, handle them gently and avoid using a pipette to mix by blowing.
• It is generally recommended to transform only a portion of the samples. In case of transformation failure, there will still be samples available for another attempt.
• All components of this kit have been sterilized. Please operate in a clean area and seal the kit after use to avoid contamination.
• This product is intended for scientific research by professionals only and should not be used for clinical diagnosis or treatment, or in food or pharmaceuticals. Do not store in a regular household.
• For your safety and health, wear lab coats and disposable gloves when handling.

Related:

• Yeast Colony PCR Kit (Alkaline Lysis)
• Yeast Colony PCR Kit (Enzymatic Lysis)
• Competent Cell Preparation and Transformation Kit for Pichia Pastoris
• Competent Cell Preparation and Transformation Kit for Saccharomyces Cerevisiae

Only for research and not intended for treatment of humans or animals

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SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory