PichiaPink™ Media Kit
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Invitrogen™

PichiaPink™ Media Kit

The PichiaPink™ Media Kit is a set of convenient prepackaged media pouches for use with the PichiaPink™ Yeast Expression System.Read more
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Catalog number A11156
Price (USD)
1,128.00
Each
Add to cart
Price (USD)
1,128.00
Each
Add to cart
The PichiaPink™ Media Kit is a set of convenient prepackaged media pouches for use with the PichiaPink™ Yeast Expression System. The PichiaPink™ Media Kit contains media needed to grow Pichia pastoris strains from frozen stocks to cultures ready for transformation and selection. The PichiaPink™ Media Kit is an easy way get started using Pichia pastoris for protein production. Use the PichiaPink™ Media Kit to refill your PichiaPink™ Yeast Expression System Kits.

The PichiaPink™ Media Kit comes with prepackaged pouches for the following solutions. Each pouch makes 1 liter of solution. Media pouches should be kept dry and stored at room temperature.

MediaUsePouches
YP+agarFor plates to streak out colonies2 pouches
YPFor liquid cultures2 pouches
PAD (Pichia Adenine Dropout)+agarFor plates to grow PichiaPink™ recombinants2 pouches
YP+sorbital (YPS)For recovering PichiaPink™ transformants2 pouches
DextroseFor adding to YP and YP+agar1 pouch

Why Choose the PichiaPink™ Yeast Expression System?
The PichiaPink™ Yeast Expression System is based on the yeast Pichia pastoris. Advantages of Pichia pastoris include rapid growth, well-defined genetic background, simple media formulation, and easy handling. For over 30 years, Pichia pastoris has been used by labs around the world for producing hundreds of different proteins from many species including human (Ref 1, 2). The PichiaPink™ Yeast Expression System allows convenient and cost-effective protein production from small to large scales.

For information on obtaining a commercial-use license for the PichiaPink™Yeast Expression System, please inquire at outlicensing@lifetech.com.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

Related Links
• Learn more about the PichiaPink™ Yeast Expression System.
• Learn more about our other protein expression systems.

References

1. Cereghino JL, Cregg JM. Heterologous protein expression in the methylotrophic yeast Pichia pastoris. FEMS Microbiol Rev. 2000 Jan;24(1):45-66. [PubMed]
2. Cereghino GP, Cereghino JL, Ilgen C, Cregg JM. Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris. Curr Opin Biotechnol. 2002 Aug;13(4):329-32. [PubMed]
For Research Use Only. Not for use in diagnostic procedures.
Specifications
FormPowder
FormatKit
Media TypePAD, YP, YPS
Preparation MethodAutoclave
Target Organism ClassP. pastoris
Final Product TypeLiquid Medium, Agar Plates
For Use With (Equipment)A11150, A11151, A11152, A11153
Quantity1 Kit
Shipping ConditionRoom Temperature
Strain DesignationsPichiaPink™
Unit SizeEach
Contents & Storage

Contains pouches sufficient to make 4 basic media:

2 pouches PAD agar, 2 pouches YPDS, 2 pouches YPD, 2 pouches YPD agar, 1 pouch Dextrose


Store at room temperature

Frequently asked questions (FAQs)

I inoculated 25 mL of MGY with one colony and grew it overnight, but the OD is not 2-6 as mentioned in your manual. Is there something wrong?

This is likely due to low inoculum. We recommend having a starter culture to use, as it is more accurate than using a colony. You can use the doubling time to calculate the starting OD. In YPD, the doubling time is about 1.5 hours. In MGY, the doubling time is going to be about 3 hours.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

When selecting for blasticidin-resistant transformants in the X-33 strain using pPIC6/pPIC6α vectors, why do I get large and small colonies on YPD plates containing 300 µg/ml blasticidin?

Generally, large colonies represent transformants containing pPIC6/pPIC6α integrants, while small colonies represent transformants containing pPIC6/pPIC6α non-integrants. These non-integrants have transduced the pPIC6/pPIC6α plasmid, and therefore, exhibit a low level of blasticidin resistance in the initial selection process. Upon subsequent screening, these non-integrant transformants do not retain blasticidin resistance.

When choosing a blasticidin-resistant transformant for your expression studies, we recommend that you pick blasticidin-resistant colonies from the initial transformation plate and streak them on a second YPD plate containing the appropriate concentration of blasticidin. Select transformants that remain blasticidin-resistant for further studies.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

My transformation is not working. Do you have any suggestions?

Here are some suggestinos:

- Make sure that you have harvested cells during log-phase growth (OD <1.0 generally).
- If electroporation is being used, see the electroporator manual for suggested conditions. Vary electroporation parameters if necessary.
- Use more DNA.
- Use freshly made competent cells.
- If the LiCl transformation method is being used, try boiling the carrier DNA.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

My spheroplasting of Pichia worked twice, but hasn't worked since. The OD of the culture simply does not drop.

Here are some things to consider:

- If the OD of cells that are used is too high, they will not spheroplast. Do not overgrow cells.
- Do not use old cells and make sure that they are in log phase of growth.
- Make sure to mix zymolyase well before using. Zymolyase is more of a suspension than a solution.
- Make the PEG solution fresh each time and check the pH.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

Is there a functional difference between using peptone or tryptone for Pichia media?

Bacto-Tryptone and Bacto-Peptone are two different and specific types of peptones. Bacto-Tryptone is a slightly poorer nitrogen source, and more of the nitrogen is provided by tyrosine and tryptophan. When comparing the two as components of media for Pichia growth, growth curves may differ slightly, but there should be only minor differences between the two. In Pichia media formulations that include Yeast Nitrogen Base as a primary source of nitrogen, such as BMGY and BMMY, there should be very little or no difference.

Find additional tips, troubleshooting help, and resources within our Protein Expression Support Center.

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