Table 2.
DICER1 oligonucleotide primers used in this study
| Exon | Forward | Reverse | Size of fragment | Annealing temp. Sanger sequencing | Annealing temp. HRM |
|---|---|---|---|---|---|
| 2 |
GCAATGAAAGAAACACTGGATG |
TCAAATCCAATTACCCAGCAG |
358 |
[1] |
64 |
| 3 |
TTTTGTAAATTTATTGGAGGACG |
TCTGCCAGAAGAGATTAAATGAG |
429 |
[1] |
64 |
| 4 |
TTTTGGAGGATAACCTTGGAAC |
AAATCAGACAACCAAGGCTACAG |
390 |
[1] |
66 |
| 5 |
TTGTCGTCAAGACATGCTTTC |
TTTAATATTCATTCATTCATACACTGC |
518 |
[1] |
66 |
| 6 |
TAGTGGCATTTCCACCAAAC |
GAATTCTTACTCTTGCCCATTCC |
437 |
[1] |
69 |
| 7 |
TTCTCACTACTGCAGTATTGATACCTT |
GAGCCGCATTAAGCATATTTTC |
303 |
[1] (7 F modified) |
69 |
| 8 |
AAATCCCAGTTAAACCCCAC |
TCACATCACAACACAGGACG |
554 |
[1] |
68 |
| 9 |
TAAATCACCGTCGCCAAATC |
AAATCACTCTACAGCTACCTCATGG |
591 |
[1] |
69 |
| 10 |
CATGTGTGTCAGAAATGACAGTTG |
TTCCTATGGATACAAAGAATAACAAAG |
431 |
[1] |
68 |
| 11 |
AGCAGGTTACTTTGGAGTACTGAAG |
AACTTTTATTGCTGCACGATACTG |
498 |
[1] |
69 |
| 12 |
TCACATTTCAAGTGCTCACC |
TGAACATGTAGATGACTACAAAAGC |
596 |
[1] |
69 |
| 13 |
TTTTACTAGGCAGGACTTTTAAAGATG |
AAGTGTTCATGGTGCATGATTC |
585 |
[1] |
NA |
| 14 |
TTTGCAGTCCAGCTCATATTG |
AAGCTGTGAATCGGAGAAAG |
498 |
[1] |
69 |
| 15 |
TAAGAAGTGTCATGCCTCGG |
TCTAGTGGAGAAATAGAAGAGGCAC |
468 |
[1] |
68 |
| 16 |
GAAAGCATCATTTCTGTTCTGAAG |
AAGAGAAAAACGACTCTTTAGC |
443 |
16R is new |
65 |
| 17 |
TTCAGCATACTGTGTTCTACCTCTT |
TTTTAGTAGAGACGAGGTTTCACC |
484 |
17 F is new |
69 |
| 18 |
TGTAAAGGTGCCATTTAGCTTC |
TTTGTGTGCAAAGCATCTCC |
589 |
[1] |
69 |
| 19 |
ATTGCACTTGAGGGATTCTTACC |
TTTGTGATATATTAATGGGCCAAG |
496 |
[1] |
67 |
| 20 |
TTGGCCCATTAATATATCACA |
TCTCACTCCAACTGTTATGGCTTA |
594 |
[1] |
NA |
| 21-1 |
AATTGCTGTTGCTCTCAGCC |
GAGTACATTCATCGCTGGGC |
508 |
[1] |
68 |
| 21-2 |
ACAAGCAGGAAATACCCGTG |
ACTGCAAACCACTTTCAGGC |
501 |
[1] |
68 |
| 22 |
AAAGCATAGAATATGTGGGAATT |
AGAAATTTGCCTCCATCAAA |
584 |
[1] |
NA |
| 23-1 |
AACCCTTGCTTTTATTGAGTTTC |
CAGGGCTTCCACACAGTCC |
574 |
[1] |
NA |
| 23-2 |
AAACTGTGGTGTTGACACGG |
TACAAGGCCAACACGATGAG |
571 |
[1] |
68 |
| 24 |
TGTGGGGATAGTGTAAATGCTTC |
TGCCGTCAGAACTCTGAAAC |
403 |
[1] |
68 |
| 25-26 |
TGGACTGCCTGTAAAAGTGG |
TGAACTTTTCCCCTTTGATG |
450 |
[1] |
66 |
| 27 | CCTGTCTGTCGGGGGTATG | TCTGCCTTCAATTCATTCCA | 448 | [1] | 69 |
Key: HRM- High Resolution Melt. Oligonucleotide primers 16R and 17 F are new; they were designed specifically for High Resolution Melt analysis.
NA: Not Applicable – the fragments amplified were not included in the HRM assay, they were sequenced instead.
[1] refers to the source of the oligonucleotide primer sequences. bp - base pairs.