Primer sequences used
Primers . | Direction . | Exon . | Annealing temperature, °C . |
---|---|---|---|
5′-CAGCTCAAGGGACTCAGCCAA-3′ | Sense | Exon 1 | 65 |
5′-CAAATCCTCGGTTAGACCCTCTC-3′ | Antisense | Exon 1 | 65 |
5′-GAATGGAATATGAGGTCCTACTC-3′ | Sense | Exon 2 (1st segment) | 58 |
5′-AGACTCTCGAAAAAGACGTTATC-3′ | Antisense | Exon 2 (1st segment) | 58 |
5′-ACACCCGCCCTGGATAACGTCTT-3′ | Sense | Exon 2 (2nd segment) | 61 |
5′-CGCGACTCTTTGGATCTTTGCTC-3′ | Antisense | Exon 2 (2nd segment) | 61 |
5′-GTCCTAACCCAGGCCAACCT-3′ | Sense | Exon 3 (1st segment) | 61 |
5′-GGCCATCTCCTCGTAGTCGC-3′ | Antisense | Exon 3 (1st segment) | 61 |
5′-AGCTGCTGGAGGCTGTGGAC-3′ | Sense | Exon 3 (2nd segment) | 61 |
5′-CTGACTCGCCTTCCCTAGGG-3′ | Antisense | Exon 3 (2nd segment) | 61 |
5′-CAACAGGGACCTCCTCTCTATC-3′ | Sense | Exon 4 | 62 |
5′-CTGCACACACAGGGACAGCAG-3′ | Antisense | Exon 4 | 62 |
5′-GTACCTGGCCCTGGGGCTGCTG-3′ | Sense | Exon 5 | 66 |
5′-GATTCCCACCCCAGTGCCAGCT-3′ | Antisense | Exon 5 | 66 |
5′-CACTCACCCCTGCAGGCACCTGA-3′ | Sense | Exon 6 | 60 |
5′-GCACCGCCCAGACCCTGGCGG-3′ | Antisense | Exon 6 | 60 |
5′-CAGCATGCATCCTAGGCAGTTCA-3′ | Sense | Exon 7a | 65 |
5′-TCTGAGAGTCACCTGCTGTGGCT-3′ | Antisense | Exon 7a | 65 |
5′-AGGTCTCTTCCAGCCCTCAGTCT-3′ | Sense | Exon 7b (1st segment) | 61 |
5′-CCTTTGCAGCTGGGCAGGATGTT-3′ | Antisense | Exon 7b (1st segment) | 61 |
5′-AAGCCAGCACTCCTGCCATCACA-3′ | Sense | Exon 7b (2nd segment) | 61 |
5′-CTGTAAGAGCATGGGCTGGAGGA-3′ | Antisense | Exon 7b (2nd segment) | 61 |
5′-GTGTTCTGCACAGGTGCTTGC-3′ | Sense | Exon 8 | 63 |
5′-GTTGTTGGCCAGAGCTCAGAG-3′ | Antisense | Exon 8 | 63 |
5′-CACCCCAGAGCTCTCTGTGAC-3′ | Sense | Exon 9 (1st segment) | 65 |
5′-CTCGCCAGGTAGGTCTGGGCCA-3′ | Antisense | Exon 9 (1st segment) | 65 |
5′-ACTCAAGAACCTGGCCCAGACCT-3′ | Sense | Exon 9 (2nd segment) | 62 |
5′-CAGGACCTGGGAATGCTGTGATG-3′ | Antisense | Exon 9 (2nd segment) | 62 |
5′-CTGGAGGCTGTGGACGCGCGGTA-3′ | Sense | M4 | 55 |
5′-TCTTCTGGATGCTGCGGCGGAA-3′ | Antisense | R7 | 55 |
5′-AGTGTACGCCTTCTCCATCAAAG-3′ | Sense | M2 | 55 |
5′-CTTGCAGCCCTCACAGGCGCTG-3′ | Antisense | RJ | 55 |
Primers . | Direction . | Exon . | Annealing temperature, °C . |
---|---|---|---|
5′-CAGCTCAAGGGACTCAGCCAA-3′ | Sense | Exon 1 | 65 |
5′-CAAATCCTCGGTTAGACCCTCTC-3′ | Antisense | Exon 1 | 65 |
5′-GAATGGAATATGAGGTCCTACTC-3′ | Sense | Exon 2 (1st segment) | 58 |
5′-AGACTCTCGAAAAAGACGTTATC-3′ | Antisense | Exon 2 (1st segment) | 58 |
5′-ACACCCGCCCTGGATAACGTCTT-3′ | Sense | Exon 2 (2nd segment) | 61 |
5′-CGCGACTCTTTGGATCTTTGCTC-3′ | Antisense | Exon 2 (2nd segment) | 61 |
5′-GTCCTAACCCAGGCCAACCT-3′ | Sense | Exon 3 (1st segment) | 61 |
5′-GGCCATCTCCTCGTAGTCGC-3′ | Antisense | Exon 3 (1st segment) | 61 |
5′-AGCTGCTGGAGGCTGTGGAC-3′ | Sense | Exon 3 (2nd segment) | 61 |
5′-CTGACTCGCCTTCCCTAGGG-3′ | Antisense | Exon 3 (2nd segment) | 61 |
5′-CAACAGGGACCTCCTCTCTATC-3′ | Sense | Exon 4 | 62 |
5′-CTGCACACACAGGGACAGCAG-3′ | Antisense | Exon 4 | 62 |
5′-GTACCTGGCCCTGGGGCTGCTG-3′ | Sense | Exon 5 | 66 |
5′-GATTCCCACCCCAGTGCCAGCT-3′ | Antisense | Exon 5 | 66 |
5′-CACTCACCCCTGCAGGCACCTGA-3′ | Sense | Exon 6 | 60 |
5′-GCACCGCCCAGACCCTGGCGG-3′ | Antisense | Exon 6 | 60 |
5′-CAGCATGCATCCTAGGCAGTTCA-3′ | Sense | Exon 7a | 65 |
5′-TCTGAGAGTCACCTGCTGTGGCT-3′ | Antisense | Exon 7a | 65 |
5′-AGGTCTCTTCCAGCCCTCAGTCT-3′ | Sense | Exon 7b (1st segment) | 61 |
5′-CCTTTGCAGCTGGGCAGGATGTT-3′ | Antisense | Exon 7b (1st segment) | 61 |
5′-AAGCCAGCACTCCTGCCATCACA-3′ | Sense | Exon 7b (2nd segment) | 61 |
5′-CTGTAAGAGCATGGGCTGGAGGA-3′ | Antisense | Exon 7b (2nd segment) | 61 |
5′-GTGTTCTGCACAGGTGCTTGC-3′ | Sense | Exon 8 | 63 |
5′-GTTGTTGGCCAGAGCTCAGAG-3′ | Antisense | Exon 8 | 63 |
5′-CACCCCAGAGCTCTCTGTGAC-3′ | Sense | Exon 9 (1st segment) | 65 |
5′-CTCGCCAGGTAGGTCTGGGCCA-3′ | Antisense | Exon 9 (1st segment) | 65 |
5′-ACTCAAGAACCTGGCCCAGACCT-3′ | Sense | Exon 9 (2nd segment) | 62 |
5′-CAGGACCTGGGAATGCTGTGATG-3′ | Antisense | Exon 9 (2nd segment) | 62 |
5′-CTGGAGGCTGTGGACGCGCGGTA-3′ | Sense | M4 | 55 |
5′-TCTTCTGGATGCTGCGGCGGAA-3′ | Antisense | R7 | 55 |
5′-AGTGTACGCCTTCTCCATCAAAG-3′ | Sense | M2 | 55 |
5′-CTTGCAGCCCTCACAGGCGCTG-3′ | Antisense | RJ | 55 |
The table lists the sequence of the 14 sets of primers generated to amplify the PML exons. The primers were designed at the exonic-intronic boundaries to produce PCR products of various sizes, ranging from approximately 500 to 300 base pairs (bp). PCR products greater than 400 bp were digested with MboII (second fragment of exon 2 and second fragment of exon 7b) or BamHI (first fragment of exon 7b). The primers (M4, R7, M2, RJ) used to amplify and sequence the PML-RARα fusion gene are also reported.