MSSCP applications

ONCOLOGY


The discovery and detection of a minor genetic variants at the heterogenic tumor material correlating with example chemo- and radioresistance are one of the major obstacle in present anticancer therapy. The NGS based methods are very powerful tool for GWAS study but not widely adopted in routine oncology diagnostics yet. Also because of the minor genetic variants detection limits.

We have demonstrated in numerous studies (for details, please see Reference section) that the MSSCP method is an efficient technique for minor genetic variants discovery and detection at heterogenous tumor material.

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Example 1. Discovery and detection of the of the kRas gene minor genetic variants at tumor lung samples by MSSCP method

Figure 1. MSSCP electrophoretic profiles of kRas ex 1 amplicones. In samples number 3 and 4 the presence of additional ssDNA bands marked with red arrows, not visible in reference WT samples (1 and 2), was observed. Blue arrows indicate representative ssDNA bands in agreement with WT profile.

Example 2. HER2 gene minor genetic variants in metastatic lung cancer clinical samples

Figure 2. MSSCP electrophoretic profiles of ex20 HER2 samples. PCR products of ex20 HER2 gene amplification were heat denatured, and ssDNA was separated on 9% PA gel using MSSCP method under optimal electrophoretic conditions. DNA bands were visualized with silver stain. ssDNA bands corresponding to WT and mutated (MT) sequences are indicated by arrows.


Figure 2. MSSCP electrophoretic profiles of ex20 HER2 samples. PCR products of ex20 HER2 gene amplification were heat denatured, and ssDNA was separated on 9% PA gel using MSSCP method under optimal electrophoretic conditions. DNA bands were visualized with silver stain. ssDNA bands corresponding to WT and mutated (MT) sequences are indicated by arrows.


Example 3. The FLT3 gene minor genetic variants discovery at lung cancer tumor material by the MSSCP method

Figure 3. MSSCP electrophoretic profiles of Flt3 ex 14 amplicones. In samples number 1 and 2 the presence of additional ssDNA bands marked with arrows, not visible in reference WT samples (3), was observed.

MSSCP electrophoresis appeared highly efficient in detection of minor genetic variants of above mentioned genes, as well as: p53 (ex 6, 7, 8), EGFR (ex 19,20, 21), nRas (ex 1), MAPK-1 interacting protein, SMARCA4 ones.

 

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