Product FAQ

My real-time PCR run stopped before it completed. What are the possible causes?

Answer

1) Power outage and/or power fluctuation. It is recommended that your instrument be equipped with an uninterruptible power supply (UPS). Several companies manufacture such systems for scientific equipment, and they are listed in FAQ Answer ID 1935.

2) Launching other programs, setting up another run or analyzing the data while a run is in progress. The real-time data collection consumes a large amount of hardware resources. Other applications may interfere with the run and disrupt data collection causing the software to crash.

3) Hard drive fragmentation. When the SDS and other programs run, many small program files are automatically generated and deleted in the hard drive, creating blank areas between files. This can slow down your system performance by as much as 50% since programs have to access more areas of the hard drive to run. It is recommended to defragment the hard drive once a month. Please see FAQ Answer ID 2713 for more information about how to defragment a hard drive.

Answer Id: E2716

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Product FAQ

Can human PSCs be frozen in Gibco Essential 8 Medium?

Answer

Yes, cells can be routinely frozen in complete Gibco Essential 8 Medium and 10% DMSO.

Answer Id: E12444

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Product FAQ

Can I warm Gibco Essential 8 Medium in a 37 degrees C waterbath for daily use?

Answer

It is very important that complete Gibco Essential 8 Medium is prewarmed at room temperature and not in a 37 degrees C water bath. bFGF activity can decline rapidly with repeated temperature changes from 4 degrees C to 37 degrees C.

Answer Id: E12433

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Product FAQ

What is the role of insulin in Gibco Essential 8 Medium?

Answer

Insulin is important for cell survival and proliferation.

Answer Id: E12445

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Product FAQ

How many components are there in Gibco Essential 8 Medium?

Answer

Gibco Essential 8 Medium contains only the eight required components for culturing PSCs. The medium was developed by Chen et al (http://www.ncbi.nlm.nih.gov/pubmed/21478862) to overcome the variability issues observed with mTeSR Medium. Gibco Essential 8 Medium is designed to have less variability due to limited components and removal of albumin (BSA) from the formulation. Gibco Essential 8 Medium is provided as a convenient two-component kit: 500 mL Gibco Essential 8 Basal Medium and 10 mL Gibco Essential 8 Supplement (50X).

Answer Id: E12418

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Product FAQ

Can Rho-associated protein kinase (ROCK) inhibitors be used in Gibco Essential 8 Medium?

Answer

Yes. However, this isn't necessary, and we do not routinely use these inhibitors in house. If the use of a Rho-associated protein kinase (ROCK) inhibitor is desired, you should add the inhibitor only to the medium at passage. Inhibitors should not be present for routine feeding. Use of inhibitors is assay dependent and not required for routine cell culture.

Answer Id: E12434

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Product FAQ

Does Gibco Essential 8 Medium already contain bFGF?

Answer

Yes. Gibco Essential 8 Medium contains 100 ng/mL basic fibroblast growth factor (bFGF), and no additional bFGF is required.

Answer Id: E12419

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Product FAQ

What is the role of a Rho-associated protein kinase (ROCK) inhibitor or blebbistatin?

Answer

The inclusion of either a ROCK inhibitor (HA100 or Y27632) or blebbistatin improves initial survival and supports a high cloning efficiency, which is increased by the addition of transferrin and selenium. If cells are cultured routinely in medium containing a ROCK inhibitor, it may become necessary to include it for routine culture.

Answer Id: E12435

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Product FAQ

Is Gibco Essential 8 Medium xeno-free (human or recombinant origin only)?

Answer

Yes. Gibco Essential 8 Medium provides reliable and robust cultures with a xeno-free, eight-component medium

Answer Id: E12423

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Product FAQ

Does Gibco Essential 8 Medium exhibit variability from lot to lot?

Answer

Gibco Essential 8 Medium has reduced variability compared to existing feeder-free culture media. Unlike other media that contain over 20 highly variable ingredients, Gibco Essential 8 Medium is produced under cGMP and has an optimized formulation and growth factor levels that help ensure maximum cell health, pluripotency, and growth, with minimal variability.

Answer Id: E12426

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Product FAQ

Can I feed the cells grown in Gibco Essential 8 Medium less frequently than those in other feeder-free medium?

Answer

No. The cells should be fed daily including the day after passaging.

Answer Id: E12436

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Product FAQ

How do I prepare Gibco Essential 8 Medium?

Answer

To prepare 500 mL of complete Gibco Essential 8 Medium, thaw Gibco Essential 8 Supplement (50X) at room temperature for 1 hour or 2-8 degrees C overnight and then aseptically combine the components listed below:

- Gibco Essential 8 Basal Medium: 490 mL
- Gibco Essential 8 Supplement (50X): 10 mL

Answer Id: E12429

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Product FAQ

What is the shelf life of complete Gibco Essential 8 Medium?

Answer

The shelf life of complete Gibco Essential 8 Medium is two weeks at 2-8 degrees C.

Answer Id: E12432

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Product FAQ

What is Gibco Essential 8 Medium?

Answer

Gibco Essential 8 Medium is a xeno-free and feeder-free medium specially formulated for the growth and expansion of human pluripotent stem cells (PSCs). Originally developed by Chen et al (http://www.ncbi.nlm.nih.gov/pubmed/21478862) in the laboratory of James Thomson, and validated by Cellular Dynamics International, Gibco Essential 8 Medium has been extensively tested and is proven to maintain pluripotency in multiple PSC lines.

Answer Id: E12417

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Product FAQ

What are the main differences to be taken into consideration when culturing cells in Gibco Essential 8 Medium on Gibco Vitronectin (VTN-N) compared to other feeder-free systems?

Answer

Here are three major differences to be taken into consideration when culturing cells in Gibco Essential 8 Medium on Gibco Vitronectin (VTN-N) compared to other feeder-free systems:

- Cells should be typically passaged ~24 hours sooner than they would be with other feeder-free media.
- Passaging should take place when cells are at ~85% confluency. If cells are passaged when they are more than 85% confluent, the health of the cells and final cell yield may be compromised.
- Cells must be passaged in EDTA. Collagenase and dispase are not recommended.

Answer Id: E12422

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