Ultra Yield Flask FAQ

Why do Ultra Yield Flasks work better than Glass Flasks, and other Disposable baffled flasks for e.coli, Pichia, Pombe, Scizomyces?
Ultra Yield Flasks (patented) are a disposable high aeration shake flask or bioreactor. Ultra Yield Flasks achieve high aeration with a 6 baffle design that has been optimized for fermentation for e.coli, Pichia, Pombe, Scizomyces, and other microbes. They provide good airlift with high speed shaking giving an increase of 10 fold oxygenation, and when combined with nutrient enriched media and proper pH balance, can increase yield significantly. This, combined with the simple AirOtop Enhanced seal made from airporous material allows great aeration and is disposible. No cutting of aluminum foil, and exposure to cross contamination with used flask plugs. Ultra Yield Flasks allow for scalable results from the 125mL, 250mL and 500mL up to the 2.5L.
How does the yield increase with Ultra Yield Flasks?
The Ultra Yield Flasks work to grow more biomass or cells acting as a disposable bioreactor. This leads to an increased protein, DNA, etc. and with an increase in density there comes an increase in yield. Cells in Ultra Yield Flasks can be grown for more than 24 hours, because you are not oxygen limited. Cells do not go from log phase to stationary phase but as shown by data (GSK Data ) go to a transition phase. This transition stage allows for continued growth without anaerobic conditions that use the cells' own nutrients to continue its living without stressing the system. pH maintaince is extremely important so buffering with a non-temperature dependant buffer like MOPS makes a large difference in cell growth with the Ultra Yield Flasks. Ultra Yield Flasks allow for optimum growth in shake flasks, and allow for scaleable results from the 125mL flask-2.5L flask. In studies and testing we have found that Ultra Yield Flasks result in consistent standardized results.
What ratio of liquid media can we have in the Ultra Yield Flasks for e.coli and microbes?
What are the recommended speeds for e.coli and microbes?
Media ratio depends on the cells being grown. We are going to give a general rule for E.coli. E.coli use a 40% fill volume. In the 2.5L, some people put 500mL for maximum OD, and others use 1L. They use 3-4 flasks to replace a small scale fermentor.

Recommended speeds:

Recommended culture volumes and orbital shaker speeds (a throw of 1" is generally used for orbital shaking)

Part # Description Media (mL)/Flask
Shaker Speed
931147
Please cover the flask with AirOtop Enhanced seals 899421.
125mL Ultra Yield Flasks 50/case -- Sterile
40-50mL/flask 300-350 RPM
931144
Please cover the flask with AirOtop Enhanced seals 899423.
250mL Ultra Yield Flasks 50/case -- Sterile
75-110mL/flask 300-350 RPM
931141
Please cover the flask with AirOtop Enhanced seals 899424.
500mL Ultra Yield Flasks 50/case -- Sterile
125-200mL/flask 300-350 RPM
931136-B
Please cover the flask with AirOtop Enhanced seals 899425.
6/case 2.5L Ultra Yield Flasks -- Sterile
500mL (optimum)
300-400 RPM
931136-B
Please cover the flask with AirOtop Enhanced seals 899425.
6/case 2.5L (Fermentor Mode)-- Sterile 1L 250 RPM
Chart for 1" Orbit | Click Here for 2" orbit Data
Has anyone tested the Ultra Yield Flasks for acetate concentration?
Can you use it as a starter culture for Vaccines?
GSK has kindly provided data on pH, OD, and acetate on their use before Vaccines. Acetate seems to gain with normal shake flasks, whereas pH and Acetate both stay consistent with Ultra Yield flasks. The flasks actually have shown that they reduce content over time.
What clamps and shakers work best for the Ultra yield flasks?
Ultra Yield Flasks are designed to shake in 1 inch orbit shakers. With the higher speeds sticky tape will not work, as higher shaking speeds allow for better aeration. Our 125ml, 250mL and 500mL will work with standard shake flask clamps. The 2.5L can take a standard 2L shake flask clamp, and New Brunswick has a special clamp. Kuhner shakers have also made a special flask clamp.
What media do we put in the Ultra Yield Flasks?
Recommended media is an enriched media with pH balance around 7.4 . TB, EnBase, Magic Media are all good examples of highly enriched medias. The pH balance is most useful with buffers that contain phosphate since these are not temperature dependant. Personally, we prefer MOPS at 100 millimolar with any of these medias.
Are the Ultra Yield flasks disposable or re-usable?
The Ultra Yield flasks are priced to be a disposable bioreactor or disposable shake flask. Ultra Yield flasks are reusable, but the cost of the flasks is significantly cheaper than the autoclaving. With labor being around $100.00/per hour it does not make sense to reuse the product? This is why we made the Ultra Yield Flasks a few dollars. If you are to reuse the flasks we would suggest three times before any drop off in density, as we know plastic can become brittle over time. Some people have used the flasks multiple times, but remember a little detergent in the flasks and hurting an experiment is more expensive than just using a new one.
Myths and legends of growing cultures?
  1. LB is the least healthy growth media. Ultra Yield Flasks are best when using an enriched media.
  2. Growing cultures for more than 18 hours will damage my culture. What damages the growth and causes anaerobic growth is the insufficient amount of oxygen, and drop in pH when using normal shake Flasks. Ultra Yield Flasks give sufficient aeration for 7 days worth of growth at 20*C, and 50 hours at 37*C.
What is missing with LB and minimal media?
LB or minimal media is like feeding someone bread and water. One needs to get roughage, and nutrients and vitamins. This is true with other living organisms as microbes need a balanced diet to grow strong.
Media choices and recommendations for end users?
Different types exist, and by no means is this an endorsement of one over another; TB, Magic Media, Enbase, etc.
What systems are people using with Ultra Yield Flasks?
People use the shake flasks, on New Brunswick, ATR, Kuhner, and every shaking platform available.

End user Question:

We grow organisms that need to be killed after cultivation by autoclaving. Can the flasks withstand autoclaving for this purpose? Both flasks and seals are sterile and ready to use.
Ultra Yield Flasks and Seals come ready to use sterile. As described, people add media, and sterilize the flask with the media in it, but do not use the AirOtop Enhanced seal for this process. For sterilization it is best with a piece of Aluminum foil on top. After Autoclaving allow the Ultra Yield Flasks to cool down before using as they do retain heat. Customers have erred with killing their cells by taking them straight from the Autoclave to adding their cells immediately for growth.
How do you grow your cells and media?
We usually prepare a large amount of media and sterilize it in bottles. Then pour the ready made media into the Ultra Yield Flasks, and add the cells. We find this works best, and is fast for doing multiple runs. This is easier.
Applicable Notes: