[DIYbio] Re: Any yeast culturing and/or gen transfer protocol, simple enough to do at a simple laboratory and sourcing yeast?

Hi Shamrock

Thanks for the comprehensive and informative answer. Cheers!

On 6/24/12, shamrock <thmsburkett@gmail.com> wrote:
>
>
> Hi Brian,
>
> Bakers yeast (Saccharomyces cerevisiae) is just as easy to work with as E.
> coli, keeping in mind a few things. One minor complication is that they
> grow slower and most antibiotics don't work as selections. What that means
> from a practical standpoint is that contamination becomes a slightly
> greater problem then when working with E. coli, and the media is a bit more
>
> complex to make. However if you keep a clean environment and use good
> aseptic techniques you shouldn't have much of a problem.
>
>
>
> The background yeast strain used for most genetic experiments is one known
> as S288C. Other strains and backgrounds are used for a variety of
> industrial, brewing and baking purposes but S288C is the one most often
> used in genetic experiments. It has the advantage that it does not have a
> lot of chromosomal duplications and gene amplifications that are found in
> most industrial strains, plus a wide variety of different strain types are
> avaiilable.
>
>
>
> There are a number of repositories that have collections of yeast
> strains-alas I do not know how willing they would be to send something to a
>
> home biologist. You could search on the internet for biological
> repositories or if you have access the latest issue of journal of
> industrial microbiology and biotechnology has a review on yeast culture
> collections (Boundy-Mills, K. 2012. Yeast culture collections of the world:
>
> meeting the needs of industrial researchers. JIMB 39:673).
>
>
>
> Like I said earlier working with yeast is just as easy as E. coli with a
> few caveats. First of all because most antibiotics don't work the most
> often used selections are nutritional selections. What this means is that
> the starting strain is unable to make a particular nutrient (usually an
> amino acid or a nucleotide precursor) because of a mutation in a single
> gene. The selectable marker carried on the plasmid has an unmutated copy of
>
> the same gene. Therefore cells carrying the plasmid can grow in the absence
>
> of the nutrient but cells that do not have the plasmid cannot-just like E.
> coli cells with a plasmid can grow in the presence of ampicillin but cells
> that don't have the plasmid cannot. From a practical standpoint it means
> that the media used is a bit more complex so instead of growing cells in
> rich media (which is relatively simple) you need to select for recombinant
> cells on media that lacks only that single nutrient. You can but the media
> known as drop-out media or you can make it yourself but it is more complex
> to make from scratch. Cells are transformed similar to E. coli cells
> although the method used to make the cells competent to take up DNA is
> different. With yeast Li salts (Li acetate for cerevisiae and LiCl for
> Pichia ) are used. You can also electroporate yeast like you can E. coli,
> and there is a method based on digesting away the outer cell wall.
>
> A number of different plasmid vectors exist for use in yeast and a number
> of different inducible promoters are available as well. So you can make use
>
> of high oor low copy episomal vectors or you can use integrating vectors.
> Promoters, such as the GAL promoter, can be turned on just by shifting the
> carbon source from glucose to galactose (making sure the glucose is used
> up). One really cool thing about yeast is that it is relatively easy to
> make genome modifications either by knocking out certain genes or adding
> new ones. Plus you can do some nice genetic experiments through simple
> mating and sporulation.
>
> Working with yeast is fun, pretty easy to do, and they smell a lot better
> than E. coli!
>
> On Saturday, June 23, 2012 4:15:23 AM UTC-4, Brian Soe wrote:
>
>> Hi friends
>>
>> I want to learn some biology experiment at home for fun. I wonder
>> is there any yeast culturing protocol that can be done without a
>> research grade laboratory but a simple bench, a light microscope,
>> micro pipette and syringe, etc. May I know where are the potential
>> sources where can I get them? Thanks.
>>
>> Sincerely
>>
>> Aung
>>
>

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