Table 1.
PCR Conditions of Individual Fragments within Intron 1 of RET Proto-Oncogene[Note]
|
Primers(5′→3′) |
|||||
| Fragment | Forward | Reverse | Size(bp) | AnnealingTemperature(°C ) | Cycles |
| P1 | M13F*-GTGGAAGTTGTGGTGAGCCAAG | M13R*-GTGGGAAGTAGGGAAGTGAGTGAG | 1,057 | 67 | 35 |
| P13 | M13F-TGGCACAATCTCGGCTCACTAC | M13R-CTTGGCTCACCACAACTTCCAC | 1,051 | 68 | 35 |
| P1b | M13F-ATGACTTTCCTGTAAAGTGC | M13R-GGAGTTTTTCATCTCTGTTC | 338 | 54 | 35 |
| P1a | M13F-AGATAAGATGCACGGACCTTAG | M13R-GCAACAGTTGCCAAAAAATG | 596 | 60 | 35 |
| P13b | M13F-CCAGAAGTGGGATTGGTAG | M13R-TCACCACAACTTCCACCTC | 597 | 59 | 35 |
| P13a | M13F-GTGCAATGGCACAATCTC | M13R-GAAAAACAAGAAGGTAGTTCCC | 552 | 58 | 35 |
Note.— Primers were designed using the PRIME command of GCG-Wisconsin software, and all of them had universal M13 primers attached at the 5′ ends. M13F*: CGCCAGGGTTTTCCCAGTCACGAC; M13R*: TTTCACACAGGAAACAGCTATGAC. Fragments P1 and P13 were each divided into two smaller fragments (a and b), to obtain better results in the performance of EMD technology. Fragments were overlapping, except for P1a and P13b, which were separated by the STR (TAAAn).