GITC——Key Reagent for Nucleic Acid Extraction and Sample Protection

发布于 2025年6月23日更新时间 2025年6月23日

Product Manager: Harrison Michael

GITC is a strong denaturant and nuclease inhibitor. It is commonly used in the form of high-concentration solutions for life science research such as nucleic acid extraction, sample preservation, virus inactivation, protein denaturation, and pre-processing of molecular diagnostics. Aladdin provides high-quality GITC isothiocyanate reagents. This article will comprehensively sort out various experimental applications of GITC starting from its chemical properties, and technically expand on key experimental processes to help researchers accurately select and use this reagent.

1. Application Advantages Determined by Chemical Properties

GITC is a white crystalline solid with the molecular formula CH₅N₃·HSCN and a molecular weight of 118.16. It has the following chemical characteristics:

· Extremely strong protein denaturation ability: It can destroy the internal hydrogen bonds of proteins and inactivate proteins.

· Efficient RNA enzyme and DNA enzyme inhibitory activity: Inhibits nucleic acid degradation.

· Both cytolytic and lysing abilities: Destroys cell membranes/viral capsids and releases cellular contents.

· Good sample stability: When combined with other reagents, it can protect nucleic acids from degradation at room temperature.

It is these characteristics that make it play an irreplaceable role in a variety of life science experiments.

2. Product Attributes

Molecular formula: CH5N3HSCN
Molecular weight: 118.160 g/mol

CAS: 593-84-0

Solubility: Soluble in water and ethanol

Density: 1.103

Sensitivity: Hygroscopic and light-sensitive

Melting point: 118-122°C

Biochemical mechanism: A potent protein denaturant that can denature endogenous ribonucleases and play a role in separating ribosomal ribonucleic acid (rRNA) from ribosomes.

Applications: As a chaotropic agent and strong denaturant, it is used for denaturing and lysing cells; extracting RNA and DNA.

Structural formula:

3. Summary of Main Experimental Applications of GITC Isothiocyanate

3.1 Nucleic Acid Extraction Field

Application	Brief Description
RNA extraction	Combined with phenol/chloroform to achieve high-purity RNA separation (such as the TRIzol method).
mRNA and miRNA enrichment	By optimizing centrifugation and precipitation conditions, RNAs of different lengths can be further separated.
DNA extraction	Particularly suitable for DNA extraction from samples with strong enzyme activity, such as feces, saliva, and CSF samples.
Viral nucleic acid extraction	While lysing the sample, inactivating the virus to ensure safe operation and high-quality RNA recovery.

3.2 Sample Lysis and Nucleic Acid Protection

Application	Brief Description
Tissue cell lysis	Rapidly destroy the cell membrane structure and release nucleic acids.
White blood cell lysis (whole blood)	Often used in combination with EDTA to prevent RNA degradation.
Microbial lysis (such as fecal flora)	Inactivate enzymes and lyse bacteria and fungi at the same time, which is conducive to the extraction of microbial DNA.
Pathogen lysis	Including bacteria, viruses, parasites, etc., and can be directly subjected to qPCR detection after lysis.

3.3 Sample Storage and Transportation

Application	Brief Description
RNA stabilizer component	Preserve RNA in blood, tissue, saliva, fecal samples at room temperature.
Sample transport buffer	Prepared into VTM (viral transport solution) or DNA/RNA protectant, suitable for long-distance transport.
Cryopreservation solution	Preserve the lysed RNA samples at -80°C for a long time to prevent degradation.

3.4 Nuclease Inactivation and Experimental Contamination Control

Application	Brief Description
RNase inactivation in experimental operations	Commonly used to treat experimental instruments or solutions to prevent RNA degradation.
DNA/RNA protection after sample processing	Adding GITC after sampling can inhibit enzyme activity, which is suitable for field sample collection and POCT pre-processing.

3.5 Virus Inactivation and Clinical Pre-Processing

Application	Brief Description
Inactivation before viral nucleic acid extraction	Samples such as novel coronavirus and hepatitis B virus can be inactivated immediately after collection.
Pre-processing of PCR/qPCR	It can safely operate potentially infected samples without a BSL-3 environment.
Pre-processing solution for automated nucleic acid extraction platforms	Improve extraction throughput and safety.

3.6 Protein Research and Inactivation Treatment

Application	Brief Description
Protein denaturation research	Destroy the secondary structure of proteins in SDS-PAGE or denaturing electrophoresis.
Enzyme inactivation experiment	Degrade the activity of protease/transcriptase for structural function research.
Inhibition of non-specific protein activity	Reduce background noise in a variety of immune response backgrounds.

3.7 PCR Inhibition Research and System Optimization

Application	Brief Description
Establish a PCR inhibition standard system	Use GITC to simulate the inhibitory effect of complex samples on PCR and optimize the amplification system.
Evaluate the removal ability of magnetic bead/column extraction reagents	Determine whether PCR inhibitors can be effectively removed.

4. Detailed Explanations of Key Experiments

The following are the detailed steps and key points of several high-frequency experiments:

4.1 Experiment 1: RNA Extraction by TRIzol Method

Applicable samples: Cells, animal tissues, blood, saliva, etc.

Core reagent: TRIzol (containing GITC + phenol)

Brief process:

1. Lyse the sample: Add 1 mL of TRIzol to the sample, pipette or homogenize.

2. Add chloroform and shake: Add 200 μL of chloroform(Chloroform substitute for RNA Extraction Buffer) and shake vigorously.

3. Centrifuge to separate phases: Centrifugation forms the upper layer (RNA), middle layer (DNA), and lower layer (protein).

4. Precipitate RNA with isopropanol: After RNA enters the aqueous phase, add an equal volume of isopropanol to precipitate.

5. Wash and redissolve: Wash with 75% ethanol, dry, and resuspend.

4.2 Experiment 2: DNA Extraction from Fecal Samples (Metagenomics or 16S)

Brief steps:

1. Add GITC lysis solution → add glass beads and shake to lyse.

2. Add proteinase K and incubate at 37°C.

3. Centrifuge to remove debris → extract DNA from the supernatant (can be combined with magnetic beads).

4. Perform library construction after purification.

4.3 Experiment 3: Viral Sample Extraction and Inactivation

Application scenario: Virus inactivation after collection of nasal swabs and saliva samples.

Operation key points:

1. The virus collection solution contains high-concentration GITC.

2. Inactivate immediately after adding the sample → reduce the risk of infection.

3. Extract high-quality RNA by subsequent magnetic bead/column method → perform RT-qPCR analysis.

5. Recommended Formula Reference (Common GITC Lysis Solution)

Component	Concentration	Function
GITC	4-6 M	Main denaturant/enzyme inhibitor.
Tris-HCl	25-50 mM	Buffer to stabilize pH.
EDTA	10 mM	Chelates metal ions and inhibits the activity of metal enzymes.
β-Mercaptoethanol	1%	Degrades redox bridges and assists in RNase inhibition.

6. Conclusion

GITC, with its protein denaturation ability, nuclease inhibition effect, broad-spectrum sample compatibility, and stability protection, runs through almost every key link in nucleic acid research, molecular diagnostics, microbial detection, and clinical sample processing. Whether researchers pursue extreme purity in RNA sequencing or clinical laboratories pay attention to biosafety in the processing of novel coronavirus samples, GITC can provide stable and reliable support.

Aladdin Scientific provides high-quality GITC products, which are suitable for the whole process from RNA extraction to complex sample processing, and support experimenters to obtain high-quality data in diversified applications.

Aladdin：https://www.aladdinsci.com/

Categories: 技术文章

Tags: RNA extraction GITC DNA extraction Nucleic Acid Extraction Sample Protection Virus Inactivation Protein Denaturant Molecular Diagnosis

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