Table 1 Nested-PCR amplification primers and protocols for EGFR exon 21
|  | Primers | Sequence 5′-3′ | PCR conditions | PCR product (bp) |
|---|---|---|---|---|
External PCR amplification | External | TCAGAGCCTGGCA | 1) 95 °C, 5 min | 297 |
Forward | TGAACATGACCCTG | 61 °C, 30 s 72 °C, 30 s 3) 72 °C, 7 min2)35 cycles: | ||
|  |  | 95 °C, 45 s | ||
External | GGTCCCTGGTGTC | 61°C, 30 s | ||
|  | Reverse | AGGAAAATGCTGG | 72°C, 30 s 3) 72°C, 7 min | |
Internal PCR amplification | Internal | ATGAACTACTTG | 1) 95 °C, 5 min | 188 |
Forward | GAGGACCGT | 2)35 cycles: | ||
Internal | GAAAATGCTGGCT | 95 °C, 45 s | ||
Reverse | GACCTAAG | 60 °C, 30 s 72 °C, 30 s 3)72 °C, 7 min |