Regulatory requirements vary by product. Please see product-specific literature to determine product use.

Product FAQ

Can fluorescent protein-expressing cells be fixed?

Answer

Yes, fluorescent protein-expressing cells can be fixed using 4% paraformaldehyde in PBS for 10 min followed by one quick PBS rinse and 3 x 5 min washes with 1 mL PBS.

Answer Id: E4932

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Product FAQ

Now that Thermo Fisher Scientific is selling the Vivid Colors Aequorea victoria fluorescent protein vectors and Clontech is not, have there been any changes to the licensing requirements?

Answer

Use of our Vivid Colors fluorescent protein vectors is described under the Limited Use Label Licenses accompanying the product. For use in research at academic or government (non-profit) agencies, the purchase of the product (as a catalog item) from Thermo Fisher Scientific is sufficient; no further licenses are required. For use in research at a commercial (for profit) entity, a commercial-use license is necessary for Aequorea victoria derived proteins; this applies to EmGFP, YFP, CFP and BFP. For any questions related to licensing, please send an e-mail to outlicensing@lifetech.com

Answer Id: E4933

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Product FAQ

I left my DNA-ExpiFectamine 293 transfection complexes sit longer than 20 minutes. Will they be okay?

Answer

The best transfection efficiencies are obtained when transfection complexes are used fresh. However, they should be stable for at least an hour.

Answer Id: E9264

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Product FAQ

What Applied Biosystems reagents and instruments can I use with TaqMan SNP Genotyping Assays?

Answer

The TaqPath ProAmp Master Mix or TaqPath ProAmp Multiplex Master Mix is recommended for TaqMan SNP Genotyping Assays. Additional PCR master mixes that can be used as well, include the TaqMan Universal PCR Master Mix, No AmpErase, TaqMan Universal PCR Master Mix, TaqMan Universal Master Mix II, TaqMan genotyping Master mix, and GTXpress Master Mix. The TaqMan SNP Genotyping Assays can be used on all the real-time PCR instruments from Applied Biosystems, including 7000, 7300, 7500, 7500 Fast, 7700, StepOne, StepOnePlus, 7900HT, ViiA7, and QuantStudio systems.

Answer Id: E2592

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Product FAQ

Does EmGFP have the F64L mutation and what effect does this mutation have?

Answer

The F64L mutation in EmGFP was purported to render better folding properties to the EmGFP protein at 37 degrees C. However, in-house studies have shown that this mutation is not necessary. This mutation is present in EmGFP (and BFP) in the pRSET vectors. The EmGFP in the pcDNA6.2 DEST vectors, pcDNA6.2 GW/TOPO vectors and plenti6.2 GW vector does not contain this mutation.

Answer Id: E4935

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Product FAQ

Is MuSeek compatible with RNA library preparation workflow?

Answer

We do not supply specific MuSeek kits for NGS library preparation from RNA samples. However, MuSeek is compatible with double-stranded cDNA substrate.

Answer Id: E8877

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Product FAQ

I am getting very poor yield of protein using the Expi293 Expression System. Do you have any suggestions for optimizing protein expression?

Answer

Please refer to the Application Note (http://www.thermofisher.com/content/dam/LifeTech/migration/files/proteins-expression-isolation-analysis/pdfs.par.61153.file.dat/expi293-protein-yield-co25763.pdf for tips on optimizing protein yield using the Expi293 Expression System.

Answer Id: E9265

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Product FAQ

The LCD on the 433A has an error "PSH". The synthesis seems to be complete. What caused this, and what needs to be done?

Answer

PSH or psh means that the barcode reader (BCR) has detected the code "all bars white". This indicates that the spring-driven device, or "Pusher" (which advances amino acid cartridges) is in position in front of the diodes. If the diodes "read" the bar on the back of the pusher, it triggers an error. This reading will stop synthesis even when "no" is chosen for BCR interrupt. It is a safety feature, assuming that no cartridge is present and if synthesis continued reagent would spill at the needle position.

If the synthesis actually is complete (all amino acids coupled), then the likely cause of this error is a mistake in the building of the "Run" file. For example, if you plan to make a 10-mer, but use a pre-loaded resin, then you will only couple 9 additional amino acids. If you were to make the same peptide sequence, but with an amide resin, then you need to couple 10 amino acids. If you were planning to synthesize a 10-mer on a pre-loaded resin, but inadvertently chose "amide resin" on the CAL (calculation) page of the Run file, then the 433A checks for amino acid #10 to add. Since there is no tenth amino acid, it does not recognize that the run is complete. It pauses with the error that the pusher is in the position of the tenth amino acid. Note that this error cannot occur if you had selected "YES" to the BCR reader interrupt feature.  Instead, the run would stop, with an error, as soon as the first erroneous amino acid cartridge was detected (usually that is the first cartridge, with the above error scenario).

Answer Id: E2593

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Product FAQ

What is the length of the well in the 1-well gels?

Answer

The length of the well in the Invitrogen 1-well gels is 7.6 cm or 2.956 inches.

Answer Id: E4963

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Product FAQ

How can I obtain the sequence of a duplex from the Stealth RNAi Human Kinase Collection?

Answer

Sequence information for the Stealth RNAi Human Kinase Collection can be provided upon request. Also, when you re-order a Stealth RNAi duplex from the collection you will receive the sequence information with the product documentation.

Stealth RNAi duplexes of interest are available for reorder at a scale of 20 nmole or higher.
1. To re-order Stealth RNAi, go to https://rnaidesigner.invitrogen.com/rnaiexpress/quickOrder.do.
2. Enter the HSS (Human Stealth Select RNAi ) or VHS (Validated Human Stealth Select RNAi) number in the field. (To obtain the HSS or VHS number of your specific Stealth RNAi, refer to the spreadsheet included in the CD-ROM.)

Answer Id: E4938

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Product FAQ

Can I use lower than recommended sample amount for MuSeek fragmentation reaction?

Answer

No, the recommended sample DNA input should be followed strictly. The MuSeek fragmentation reaction is optimized to yield optimal fragment library lengths at recommended input.

Answer Id: E8878

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Product FAQ

I am trying to use the Expi293 Expression System in 96-well microtiter plates and am not getting good yields. Do you have any tips for optimizing in this format?

Answer

Please see the Application Note (http://www.thermofisher.com/content/dam/LifeTech/migration/files/proteins-expression-isolation-analysis/pdfs.par.28891.file.dat/expi293-microtiter-plates-co25793.pdf) for tips on optimizing protein yield using the Expi293 Expression System in 96-well microtiter plates.

Answer Id: E9266

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Product FAQ

Is there any label on the "Blank Cartridge" for the 433A peptide synthesizer (PN 400198) and if not, what happens at the bar code reader?

Answer

The blank cartridges are labeled but the label does not include the name of an amino acid. They do have a bar code. It will be read as "#29", but you may, through the Software "Dictionary", change the code assignment to correctly reflect the amino acid you are using. Any amino acid cartridge without a barcode will be interpreted as though the "pusher" has reached the position before the needles. This will stop the synthesis, as continuing could mean spraying liquid from the needle onto the instrument guideway and other surfaces. This is a safety over-ride, and occurs with either "yes" or "no" selected for the barcode reader interrupt feature.

Answer Id: E2594

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Product FAQ

In the appendix section of ZOOM IEF Fractionator manual that details making the 1.1X IEF Denaturant, it states:

Filter the supernatant using a 0.2 µm filter. Use polyvinylidene fluoride (PVDF) or nylon filters. Do not use nitrocellulose or cellulose acetate filters.

Why can't nitrocellulose be used?

Answer

Nitrocellulose filters can be used, but they do contain certain charged chemicals that may seep into the buffer and may contribute to high conductivity. PVDF does not have this issue.

Answer Id: E4964

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Product FAQ

Do I need to replace the buffer every 2 - 3 days if the Applied Biosystems 3130/3130xl Genetic Analyzer is not in use?

Answer

If the instrument is not in use, it is not necessary to replace the buffer every 2 - 3 days. However, the buffer level needs to remain at the fill line to prevent the capillary or array from drying out. Top off the buffer volume with water if necessary. Replace the anode and cathode buffer with freshly made 1X Running Buffer prior to starting a new run.

Answer Id: E12697

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