KRAS Mutation Detection in Paired Frozen and Formalin-Fixed Paraffin-Embedded (FFPE) Colorectal Cancer Tissues
Abstract
:1. Introduction
2. Results and Discussion
3. Experimental Section
3.1. Tissue Samples and Processing
3.2. DNA Isolation
3.3. PCR Amplification and DNA Sequencing
3.4. HRM Analysis
4. Conclusions
References
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Nucleotide Change | Amino Acid Change | Number of Cases |
---|---|---|
c.34G>A | p.G12S | 2 |
c.34G>C | p.G12R | 1 |
c.34G>T | p.G12C | 1 |
c.35_36GT>TC | p.G12V | 1 |
c.35G>A | p.G12D | 15 |
c.35G>C | p.G12A | 3 |
c.35G>T | p.G12V | 13 |
c.37G>C | p.G13R | 1 |
c.37G>T | p.G13C | 1 |
c.38G>A | p.G13D | 8 |
c.40G>A | p.V14I | 1 |
Total | 47 |
Sample no. | Frozen | FFPE | |||||
---|---|---|---|---|---|---|---|
Direct sequencing | HRM | Direct sequencing | HRM | ||||
Nucleotide change | Amino acid change | Visual PCR band | Nucleotide change | Amino acid change | |||
8 | c.35G>T | G12V | mutation | + | c.37G>T | G13C | mutation |
11 | c.35G>T | G12V | mutation | + | c.35G>T | G12V | mutation |
13 | c.35G>A | G12D | mutation | + | c.35G>A | G12D | mutation |
18 | c.34G>C | G12A | mutation | - | - | NA | mutation |
21 | c.35G>A | G12D | mutation | - | - | NA | mutation |
24 | c.35G>T | G12V | mutation | + | c.35G>T | G12V | mutation |
25 | c.35G>T | G12V | mutation | + | - | WT | NA |
28 | c.38G>A | G13D | mutation | + | - | WT | WT |
29 | c.35G>A | G12D | mutation | + | c.35G>T | G12V | mutation |
32 | c.35G>A | G12D | mutation | + | c.35G>A | G12D | mutation |
36 | c.35G>A | G12D | mutation | + | - | WT | mutation |
38 | c.34G>T | G12C | mutation | + | - | WT | mutation |
42 | c.35G>T | G12V | mutation | + | c.35G>T | G12V | mutation |
43 | c.35G>A | G12D | mutation | + | c.35G>A | G12D | mutation |
65 | c.34G>A | G12S | mutation | + | c.34G>A | G12S | mutation |
68 | c.38G>A | G13D | mutation | + | c.38G>A | G13D | mutation |
74 | c.38G>A | G13D | mutation | + | - | WT | mutation |
79 | c.35G>A | G12D | mutation | + | c.35G>A | G12D | mutation |
80 | c.35G>A | G12D | mutation | + | c.35G>A | G12D | mutation |
84 | c.35G>T | G12V | mutation | + | c.35G>T | G12V | mutation |
91 | c.35G>T | G12V | mutation | + | c.35G>T | G12V | mutation |
96 | c.37G>T | G13C | mutation | + | - | WT | NA |
98 | c.38G>A | G13D | mutation | + | c.38G>A | G13D | mutation |
106 | c.38G>A | G13D | mutation | + | c.38G>A | G13D | mutation |
107 | c.35G>A | G12D | mutation | + | c.35G>A | G12D | mutation |
110 | c.35G>A | G12D | mutation | + | c.35G>A | G12D | mutation |
115 | c.35G>T | G12V | mutation | + | c.35G>T | G12V | mutation |
118 | c.35G>T | G12V | mutation | + | c.35G>T | G12V | mutation |
122 | c.34G>C | G12A | mutation | + | c.34G>C | G12A | mutation |
125 | c.35_36GT>TC | G12V | mutation | + | c.35_36GT>TC | G12V | WT |
127 | c.35G>T | G12V | mutation | + | c.35G>A | G12D | mutation |
129 | c.35G>A | G12D | mutation | + | c.35G>A | G12D | mutation |
131 | c.35G>A | G12D | mutation | + | c.35G>A | G12D | mutation |
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Solassol, J.; Ramos, J.; Crapez, E.; Saifi, M.; Mangé, A.; Vianès, E.; Lamy, P.-J.; Costes, V.; Maudelonde, T. KRAS Mutation Detection in Paired Frozen and Formalin-Fixed Paraffin-Embedded (FFPE) Colorectal Cancer Tissues. Int. J. Mol. Sci. 2011, 12, 3191-3204. https://doi.org/10.3390/ijms12053191
Solassol J, Ramos J, Crapez E, Saifi M, Mangé A, Vianès E, Lamy P-J, Costes V, Maudelonde T. KRAS Mutation Detection in Paired Frozen and Formalin-Fixed Paraffin-Embedded (FFPE) Colorectal Cancer Tissues. International Journal of Molecular Sciences. 2011; 12(5):3191-3204. https://doi.org/10.3390/ijms12053191
Chicago/Turabian StyleSolassol, Jérome, Jeanne Ramos, Evelyne Crapez, Majda Saifi, Alain Mangé, Evelyne Vianès, Pierre-Jean Lamy, Valérie Costes, and Thierry Maudelonde. 2011. "KRAS Mutation Detection in Paired Frozen and Formalin-Fixed Paraffin-Embedded (FFPE) Colorectal Cancer Tissues" International Journal of Molecular Sciences 12, no. 5: 3191-3204. https://doi.org/10.3390/ijms12053191