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How do I reconstitute my lyophilized oligonucleotide? How long can I store the oligonucleotide? Product FAQ

Answer

Centrifuge the tube for a few seconds to collect the oligonucleotide at the bottom of the tube. Carefully open the tube, and dissolve the oligonucleotide in the appropriate volume of TE (10 mM Tris-HCl, pH 8.0, 1 mM EDTA). TE is recommended over deionized water since the pH of water is often slightly acidic and can cause hydrolysis of the oligonucleotide. It is also best to pipette the solution up and down at least 10 times. Please visit this webpage (https://www.thermofisher.com/us/en/home/life-science/oligonucleotides-primers-probes-genes/custom-dna-oligos/oligo-technical-resources/oligo-protocols.html) for more information on how to calculate primer concentration and resuspension volume.

The lyophilized oligonucleotide is stable at -20 degrees C for 1 year or more. The oligonucleotide dissolved in TE is stable for at least 6 months at -20 degrees C or 4 degrees C. Oligonucleotides dissolved in water are stable for at least 6 months at -20 degrees C. Do not store oligonucleotides in water at 4 degrees C.

Answer Id: E2937

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What is the protocol for crosslinking IgG to Dynabeads from the Dynabeads Protein G Immunoprecipitation Kit (Cat. No. 10007D) using DSS (disuccinimidyl suberate)? Product FAQ

Answer

For crosslinking the antibody to Protein G Dynabeads, please refer to the crosslinking protocol with BS3 (bis(sulfosuccinimidyl)suberate) or Sulpha-DSS, which is an analogue of DSS. The only difference between these two molecules is that DSS is water insoluble and needs to be solubilized in DMSO before use. You can find the protocol at the following link.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center

Answer Id: E22288

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I would like to order a TaqMan SNP Genotyping Assay. Does the kit include everything I need to perform my experiment or do I need to purchase reagents separately? Product FAQ

Answer

Each predesigned TaqMan SNP Genotyping Assay is delivered in a single tube consisting of two differentially labeled allele-specific TaqMan MGB (minor groove binding) probes and a pair of PCR primers. For more information, please see the following link: https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/real-time-pcr-applications/genetic-variation-analysis-using-real-time/snp-genotyping-with-real-time-pcr.html

Besides the TaqMan SNP Genotyping Assay, you would need to purchase a Genotyping Master Mix and nuclease-free water. A list of compatible master mixes can be found in the following link: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0009593_TaqManSNP_UG.pdf.

Find additional tips, troubleshooting help, and resources within our Gene Expression Analysis & Genotyping Support Center.

Answer Id: E21528

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Can I use TRIzol Reagent to isolate RNA from a sucrose gradient fraction? Product FAQ

Answer

Yes you can. Here is a reference to a paper, as well as a brief description of method from the paper.

Genes to Cells (2001) 6:121-129 (under the heading 'RNA isolation and RT-PCR')

TRIzol LS Reagent (LifeSciences) was used according to the manufacturer's instructions to extract total RNA from sucrose gradient fractions. Briefly, 250 mL of each fraction was added to 750 mL TRIzol LS Reagent and shaken vigorously for 15 s. After a 10-min incubation at room temperature, 150 mL chloroform was added, followed by vigorous shaking and brief incubation at room temperature. Samples were then spun at 14,000 g for 10 min in a tabletop microcentrifuge. Five micrograms of nuclease-free glycogen were added to 300 mL of the aqueous phase and nucleic acids were precipitated with the addition of an equal volume of 2-propanol. After centrifugation at 14,000 g for 30 min at room temperature, the pellet was washed once with 75% ethanol and resuspended in 20 mL of nuclease-free, sterile water. Five microlitres of total RNA were used as substrate for random-primed cDNA synthesis using Superscript II modified MMLV reverse transcriptase (Gibco/Life Sciences).

Answer Id: E5229

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I isolated RNA from FFPE tissue and got very poor RNA quality and yield. How can I improve the overall RNA quality and yield? Product FAQ

Answer

These are our recommendations:

1. Upstream tissue procurement and tissue specimen preparation—if possible, fix tissues within one hour of surgical resection. Extensive degradation of RNA can occur before completion of the fixation process. The optimal fixation time is 12-24 hours, using neutral-buffered formalin or paraformaldehyde. Fixed tissues should be thoroughly dehydrated prior to the embedding process.
2. Block storage—storage of blocks without cut faces, when possible, prevents ongoing damage from exposure to atmospheric oxygen, water, and other environmental factors such as light and infestation (fungus, insects, etc.).
3. Choice of tissue type, size, and amount being used for RNA isolation—the recommended tissue thickness is 10-20 µm. The number of sections used is determined by the tissue type (which impacts cell density) and surface area (recommended size: 50-300 mm2). Excess starting material can cause filter clogging, resulting in poor yield.
4. Avoid using an excessive amount of paraffin for embedding tissues—when possible, excess paraffin should be trimmed away prior to starting the purification protocol. For xylene-based purification methods, two xylene treatments at room temperature should be sufficient for complete deparaffinization. If desired, you can perform a more rigorous 37-55 degrees C treatment for up to 30 minutes. After the xylene deparaffinization, it is crucial that the 100% ethanol is completely removed and the pellets are dry after the two 100% ethanol washes. The magnetic bead method employs novel chemistries to deal with the paraffin that limits input to 20 µm sections.

Read more about RNA isolation from FFPE tissues here (https://www.thermofisher.com/us/en/home/life-science/dna-rna-purification-analysis/rna-extraction/rna-sample-extraction/working-with-ffpe-samples.html).

Answer Id: E7868

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What are Gibco Cell Culture Bags? Product FAQ

Answer

Gibco Cell Culture Bags are convenient, pre-filled bags in a variety of formulations and sizes, specifically designed to increase productivity and reduce costs. The bags can be formatted up to 1,000 L. Read more about these bags here (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/classical-media/gibco-cell-culture-bags.html).

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

Answer Id: E11839

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What is the Gibco PE Film? Product FAQ

Answer

Based on feedback from customers, we designed a media bag portfolio that uses a consistent film for all bag sizes. This film, called PE film, is superior in function to other commercially available films. Please go here to read more about the PE Film specifications (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/classical-media/gibco-cell-culture-bags.html).

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

Answer Id: E11840

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What are the benefits of the Gibco PE film? Product FAQ

Answer

The commercially available film is a very clear, multi-layered, single web film with a polyethylene contact surface that can be made into a wide range of bag sizes (250 mL to 1,000L). In addition, the film is a market leader with exceptional gas barrier properties and very low extractable and leachable results. Click here to read more about the PE Film specifications (https://www.thermofisher.com/us/en/home/life-science/cell-culture/mammalian-cell-culture/classical-media/gibco-cell-culture-bags.html).

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

Answer Id: E11841

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Enhancement of the photoluminescence of CdSe quantum dots during long-term UV-irradiation: privilege or fault in life science research? Citations & References

  • Authors: Zhelev Z, Jose R, Nagase T, Ohba H, Bakalova R, Ishikawa M, Baba Y
  • Journal: J Photochem Photobiol B
  • PubMed ID: 15246356

Multiphoton microscopy in life sciences. Citations & References

  • Authors: König K
  • Journal: J Microsc
  • PubMed ID: 11106949
Catalog #

Fluorescence Fundamentals Molecular Probes Handbook

Blinking and nonradiant dark fraction of water-soluble quantum dots in aqueous solution. Citations & References

  • Authors: Yao J, Larson DR, Vishwasrao HD, Zipfel WR, Webb WW
  • Journal: Proc Natl Acad Sci U S A
  • PubMed ID: 16169907
Catalog #

Oligonucleotide microarrays in microbial diagnostics. Citations & References

  • Authors: Bodrossy L, Sessitsch A
  • Journal: Curr Opin Microbiol
  • PubMed ID: 15196491

Inducible Protein Expression Using the T-REx System Experiment Protocol

Inducible shRNA Expression Vectors Experiment Protocol

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