What is Guanidine Thiocyanate?

Guanidine Thiocyanate (GTC) (C833U60) is a chemical compound that is used in many RNA and DNA related research studies. Due to the emergence of COVID-19, RNA- and DNA-based viral research is crucial to help identify pathogenic viruses with epidemic potential. As the need for viral research increases, the demand for items such as GTC continues to rise.

Why is Guanidine Thiocyanate important?

As a chaotropic agent and a protein denaturant, GTC has many uses and applications. Its main functions include deactivating viruses, lysing cells and virus particles, and inactivating nucleases. In addition to these functions, GTC is also used as a storage buffer for blood samples and used to store and freeze fecal samples for DNA research.

There are many different techniques and chemicals available for these various functions. For example, urea or guanidine hydrochloride is frequently used as an alternative to GTC to inactivate nucleases. However, guanidine thiocyanate is favorable over other protein denaturants because it is a much stronger denaturant and an extreme chaotropic agent, allowing for higher yields of high-quality RNA and DNA.

GTC from MP Biomedicals/Thomas Scientific

We offer premium quality molecular grade GTC that can readily dissolve proteins, disintegrate cellular structures, and dissociate nucleoproteins from nucleic acids (due to loss of protein secondary structure), without introducing any impurities or contaminants. Available in various pack sizes and bulk quantities, our GTC provides dependability for any application from nucleic acid isolation to sample preservation.

Did you know you can make your own RNA extraction kit?

Although commercial kits are good, making your own TRIzol-based solution might be better. Not only will you save time and money, but you can also avoid long kit lead times. Producing your own solution offers increased convenience and flexibility while reducing the amount of plastic waste generated. Consider using our GTC to create your own TRIzol-based solution to isolate RNA equivalent to or better than most commercially available kits.

Make your own TRIzol-based reagent with our GTC:

TRIzol-based reagent is composed of:

• 38% Phenol
• 0.8 M Guanidine Thiocyanate (118.16 g/mol) (C833U60)
• 0.4 M Ammonium Thiocyanate (79.12 g/mol)
• 0.1 M Sodium Acetate (82.03 g/mol) (C818X57)
• 5% Glycerol (C818Z25)

1. Start with making the aqueous solution of salts and glycerol. Add the phenol when the components are completely dissolved.
2. After the solution is completely dissolved, store in a dark, cool area at around 4 °C, as the solution is light sensitive. The final pH of the solution should be in the range of 4-6.

Use your TRIZOL-based solution to isolate RNA:

1. Homogenize your sample using a homogenizer (such as MP Bio’s FastPrep^® instruments).

   1. Homogenization is different for each sample type - adjust accordingly.
      • Use 1 mL TRIzol-based reagent for every 50 mg of tissue sample
      • Use 1 mL TRIzol-based reagent for every 3.5 cm diameter culture dish (for cells in cell culture)
      • Use 1 mL TRIzol-based reagent for every 5-10 x 10⁶ animal cells used
   2. After homogenization, chill at room temperature for at least 5 minutes.
   3. Transfer the homogenate to a centrifuge to remove excess debris and transfer the supernatant (containing RNA) into a new tube.

2. Phase Separation

   1. For every 1 mL of TRIzol-based reagent used, add 0.2 mL of chloroform. For 15 to 30 seconds, shake the sample vigorously and then incubate at room temperature for a couple of minutes.
   2. Centrifuge the sample for 15 minutes and ensure the mixture is properly separated into color and colorless phases. (RNA will be in the colorless aqueous phase).
   3. Carefully remove the upper aqueous phase and place it into a separate fresh tube. (Bottom layer can be used to isolate DNA and proteins)
   4. Measure the volume.

3. RNA Precipitation and Wash

   1. Using 0.5 mL of isopropyl alcohol for every 1 mL TRIzol-based reagent used, add the isopropyl alcohol to the aqueous tube to precipitate the RNA.
   2. Incubate at room temperature for 10 minutes.
   3. Centrifuge at 12,000 x g for 8-10 minutes at 4 ºC.
   4. Remove the supernatant, and for every 1 mL of TRIzol-based reagent used, add 1 mL of 75% EtOH to wash RNA.
   5. Vortex and centrifuge again for 5-10 minutes at 7500 x g at 4 ºC. Wash once more with 75% EtOH.

4. Redissolving RNA

   1. Dry the RNA pellet briefly, but not completely, so that the solubility does not decrease.
   2. Using a pipette tip, resuspend RNA in DEPC-treated water and incubate at 55-60 ºC for 10 to 15 minutes.

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Related Products to Try

We also provide ready-to-use kits that include GTC for DNA or RNA isolation and purification. Try using one of our kits to purify your DNA or RNA:

• GENECLEAN^® Spin Kit (C837J47)
• GENECLEAN^® Turbo Kit (C837J50)
• RNAPro Solution (C831X84)
• FASTRNA^TM Pro Soil-Direct Kit (C841M38)