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What is the depth of the AlgiMatrix Matrix sponge? Will the sponge enlarge after addition of media? Product FAQ

Answer

The height of the sponge is ~3-4 mm dry. When the sponge is hydrated, its volume is essentially the same - it doesn't swell and increase in size, in fact it becomes slightly bit smaller as it essentially goes back into a hydrogel consistency, especially if the inoculated sponges are centrifuged briefly (as suggested to force cells more into the interior and also to spread the sponge uniformly over the bottom of the well).

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E11799

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Are there any additional components that need to be added to the media of the PSC Definitive Endoderm Induction Kit? Product FAQ

Answer

No other components are required.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E9380

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Are IPTG and X-gal necessary for blue/white screening of colonies? Product FAQ

Answer

With a vector containing the lac promoter and the LacZ alpha fragment (for alpha-complementation), blue/white screening can be used as a tool to select for presence of an insert. X-gal is added to the plate as a substrate for the LacZ-encoded beta-galactosidase enzyme and must always be present to see the blue color in the blue/white screening.

In cases where the lacIq repressor is present (either provided by the host cells on an F' episome, i.e. TOP10F', or expressed from the plasmid), it will repress expression of LacZ from the lac promoter and prevent the formation of the blue color with X-gal. This repression can be reversed by adding IPTG to the media in addition to X-gal, which will inhibit the action of lacIq and re-activate expression from the lac promoter.

Answer Id: E3322

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For galactose induction of expression in Saccharomyces cerevisiae, is it possible to include additional carbon sources in the media that will increase yeast growth without repressing expression from the GAL promoter? Product FAQ

Answer

Some researchers choose to grow yeast in medium containing 2% galactose as the sole carbon source during induction. However, yeast typically grow more quickly in induction medium containing 2% galactose plus 2% raffinose. Raffinose is a good carbon source for yeast, and unlike glucose, does not repress transcription from the GAL promoter. Raffinose is a trisaccharide of galactose, glucose and fructose linked in that order. Most yeasts can cleave the glucose-fructose bond, but not the galactose-glucose bond. Fructose is then used as a carbon source.

Answer Id: E3734

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For galactose induction of expression in S. cerevisiae, can I include additional carbon sources in the media to increase yeast growth without repressing expression from the GAL promoter? Product FAQ

Answer

Some researchers choose to grow yeast in medium containing 2% galactose as the sole carbon source during induction. However, yeast typically grow more quickly in induction medium containing 2% galactose plus 2% raffinose. Raffinose is a good carbon source for yeast, and unlike glucose, does not repress transcription from the GAL promoter. Raffinose is a trisaccharide of galactose, glucose, and fructose linked in that order. Most yeast can cleave the glucose-fructose bond, but not the galactose-glucose bond. Fructose is then used as a carbon source.

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

Answer Id: E9552

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Is it necessary to grow FreeStyle 293-F cells in 8% CO2 when using the FreeStyle 293 Expression Medium? If 8% CO2 cannot be arranged, can I supplement the media with an additional reagent? Product FAQ

Answer

Our recommendation of 8% CO2 for the optimal growth of Freestyle 293 cells in Freestyle 293 Expression Medium is based on the bicarbonate content in the media and growth conditions of the cells observed by R&D. That said, you may use lower concentrations of CO2 if necessary (5% is common) to grow these cells. You may observe slightly slower growth kinetics or lower cell concentrations, but functionally the entire system will still work.

Answer Id: E18338

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Can any Gibco formulation be customized with the Gibco Media Configurator? Product FAQ

Answer

Almost all Gibco cell culture basal media products can be customized using the Gibco Media Configurator. To check on a particular product, use the Media Configurator page (https://www.thermofisher.com/us/en/home/life-science/bioproduction/custom-cell-culture-media-and-services/gibco-custom-media-configurator.html to search by Gibco catalog number, or browse our offering by description. For the customization of non-media Gibco products, such as buffers and growth factors, contact us at custommedia@lifetech.com.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E11844

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What is AGT? Product FAQ

Answer

Granulated cell culture media is an easy-to-use, granular form of dry media that is produced by applying fluid bed granulation technology to cell culture nutrient media manufacturing. The patented process allows production of complete formulations of a variety of serum-free, protein-free, and chemically defined media in a dry format.

AGT offers the benefits of liquid media without the cost, storage, and transportation issues. It is a complete medium that is pH and osmolality pre-adjusted. The granules dissolve instantly for faster media preparation time than conventional dry powder media. AGT media can therefore help reduce total cycle costs, decreasing time involved in raw material planning, procurement, and testing as well as media preparation. These benefits are all realized while maintaining comparable cell growth to liquid media.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E11888

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Do I need to add additional HEPES to Hank's Balanced Salt Solution (HBSS) (Cat. No. 14025xxx)? Product FAQ

Answer

HBSS (Cat. No. 14025xxx) does not contain HEPES and it needs to be externally supplemented. The complete formulation is available here (https://www.thermofisher.com/us/en/home/technical-resources/media-formulation.153.html).

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E17448

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

Answer

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.

Answer Id: E9559

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Do you offer assistance with media formulation development? Product FAQ

Answer

Yes. Our Gibco Custom Media Specialists and R&D staff can provide advice related to the addition or elimination of media components. For more comprehensive services, our PD-Direct Bioprocess Services team specializes in media development and optimization.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E11855

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Why do FreeStyle 293-F cells require 8% CO2 for culturing? Product FAQ

Answer

The CO2 requirement is dependent on the media. Our serum-free 293 media require 8% CO2 to best buffer the media.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E17452

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Is it okay if I supplement BenchStable media with more than 10% FBS? Product FAQ

Answer

We recommended supplementing BenchStable media with at least 10% FBS for best results. If your culture method calls for greater than 10% FBS supplementation using our other existing media, we recommend that you use the same concentration with BenchStable media.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E17939

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What is the difference between Advanced RPMI 1640 Medium and other RPMI 1640 formulations? Product FAQ

Answer

Advanced Media are widely used basal media, which require 50-90% less Fetal Bovine Serum (FBS) supplementation compared to conventional basal media formulations. The Advanced media is unique due to the addition of the following ingredients to allow for serum reduction: ethanolamine, glutathione, ascorbic acid, insulin, transferrin, AlbuMAX I lipid-rich bovine serum albumin for cell culture, and the trace elements sodium selenite, ammonium metavanadate, cupric sulfate, and manganous chloride.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E13252

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What is the shelf-life of BenchStable media? Product FAQ

Answer

BenchStable media have a 12 month shelf life, consistent with our standard basal media offerings.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

Answer Id: E17926

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