ARMS-PCR For Sequence Variation Analysis

ARMS-PCR For Sequence Variation Analysis

CD Genomics is one of the professional sequencing companies with extensive experience in detecting gene mutations. We can provide high-quality ARMS-PCR service to assess sequence variations and accelerate disease research.

Amplification refractory mutation system (ARMS) is the development of the application of PCR technology. ARMS-PCR can detect two alleles simultaneously by using a single PCR reaction system, which does not require restriction endonuclease and can distinguish whether the alleles are pure or not. ARMS technology has become one of the international personalized molecular detection techniques for cancer, as well as a good method for mutation detection in genetic diseases such as sickle cell anemia and thalassemia.

The principle and process of ARMS-PCR

ARMS-PCR technology is used to detect known mutant genes. If one allele has a SNP and another alternative is normal, we can analyze the two alleles by designing specific primers for each allele. Two 5'-end primers are designed, one complementary to normal DNA and one complementary to mutant DNA. For homozygous mutations, these two primers and the 3′-end primer are added to perform two parallel PCRs. Only primers that are complementary to the mutant DNA can be extended and PCR amplified products obtained. If the mismatch is located at the 3 'end of the primer, the PCR cannot be extended.

The principle of ARMS-PCRFigure 1. The principle of ARMS-PCR. (Peng et al, 2018)

Characteristics of ARMS-PCR service

  • Fast and cheap.
  • One SNP site is detected in a single reaction.
  • Dedicated for SNP genotyping, homozygosity and heterozygosity detection.
  • The sensitivity is higher than other methods such as direct sequencing, and the mutated genes with a content as low as 0.1-1.0% in the samples can be detected.
  • In combination with real-time PCR platform, ARMS can realize closed tube operation during amplification, which is simple to operate and does not require post-processing of the product, thus avoiding the contamination of the amplification product to the greatest extent.

Workflow of ARMS-PCR service

Workflow of ARMS-PCR service

Sample requirements

  • Tissue samples: if the material provided is biological material such as fresh tissue or blood cells, please provide enough material to extract more than 2ug of genetic DNA. Stored in low-temperature refrigerator at -20℃ or -80℃, transported with liquid nitrogen or dry ice; local or short-distance transport can be ice pack transport.
  • Cultured cells in vitro (≥ 106 cells).
  • DNA samples: DNA concentration ≥ 20ng / ul, total DNA ≥ 1ug, OD260/280 between 1.7 and 2.0, without PCR inhibitor. Samples must be shipped in a cold ice pack. FFPE samples can also be received.
  • Blood samples: it is required to be stored in anticoagulant tubes or cryopreservation tubes with a volume greater than 2ml. Please transport them in dry ice to ensure that DNA is not degraded.

Want to know about other Non-NGS gene mutation detection methods?

References:

  1. Peng B, Wang Q, et al. A novel and quick PCR-based method to genotype mice with a leptin receptor mutation (db/db mice). Acta Pharmacologica Sinica, 2018, 39(1): 117.
  2. Li X, Li E, Du J, et al. BRAF mutation analysis by ARMS‐PCR refines thyroid nodule management. Clinical endocrinology, 2019.
  3. Aquino R, Protzel A, et al. Frequency of the most common mutations of the CFTR gene in peruvian patients with cystic fibrosis using the ARMS-PCR technique. Revista peruana de medicina experimental y salud publica, 2017, 34(1): 62-69.
* For Research Use Only. Not for use in diagnostic procedures.

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