Table 1.

Pairs of oligonucleotides synthesized for the MLPA technique. In all cases, universal PCR primer tags are added at 5′-end of LHS (5′-GGGTTCCCTAAGGGTTGGA-3′) and at 3′-end of RHS (5′-TCTAGATTGGATCTTGCTGGCAC-3′).

	Oligonucleotide sequences		Size (bp)
	Left hybridization sequence	Right hybridization sequence
a) PRKAR1A genea
EXON 1A	5′-GGAAAGGAGGGAGAAAAGGCAGAGGCGTC-3′	5′-AAGGGAGGCCGGAGGGAGAGTGGGGTGGA-3′	100
EXON 1B	5′-GTTTCCGGTGGAGCTGTCGCCTAGCCGCT-3′	5′-ATCGCAGAGTGGAGCGGGGCTGGGAGCAA-3′	100
EXON 2	5′-CAG GGAATACTTT GAGAGGTTGG AGAAGGTAAA-3′	5′-AATAAATGTGGGGAGATGATGAGGTGATTGTGA-3′	108
EXON 3	5′-CCAGTGGTTAAAGGTAGGAGGCGACGAGGTGCT-3′	5′-ATCAGCGCTGAGGTCTACACGGAGGAAGATGCG-3′	108
EXON 4	5′-CTAGGTTATACCAAAAGATTACAAGACAATGGCCGCT-3′	5′-TTAGCCAAAGCCATTGAAAAGAATGTGCTGTTTTCAC-3′	116
EXON 5	5′-TGCCATGTTTTCGGTCTCCTTTATCGCAGGAGAGACTGT-  GATTC-3′	5′-AGCAAGGTAAGGGCCTCTGGAGCATGCAAT-3′	116
EXON 6	5′-CTCTTTTAGGTGATGAAGGGGATAACTTCTATGTGATT-  GAT-3′	5′-CAAGGAGAGACGGATGTAAGATTTACCAATATCAAAAA-  TAT-3 ′	124
EXON 7	5′-GATTTATGGAACACCGAGAGCAGCCACTGTCAAAG-  CAAAGA-3′	5′-CAAATGTGAAATTGTGGGGCATCGACCGAGACAGCTA-  TAGA-3′	124
EXON 8	5′-CGGAAGATGTATGAGGAATTCCTTAGTAAAGTCTCTATTT-  TAGG T-3′	5′-GAGTTGTAAAGTGTGTTAACTTTGCTAGTATGTGAGA-  TACCCCTG-3′	132
EXON 9	5′-GTCTTACGGTAGCTGATGCATTGGAACCAGTGCAGTTT-  GAAGATG-3′	5′-GGCAGAAGATTGTGGTGCAGGGAGAACCAGGGGAT-  GAGTTCTTCA-3′	132
EXON 10	5′-TTGGTGATTTTATTATAGGGGTCAGCTGCTGTGCTA-  CAACGTCGGTC-3′	5′-AGAAAATGAAGAGTTTGTTGAAGTGGGAA-  GATTGGGGCCTTCTGATT-3′	136
EXON 11	5′-G TGCGTTAAGCTGGACCGACCTAGATTT-  GAACGTGTTCTTGGCCCAT-3′	5′-GCTCAGACATCCTCAAACGAAACATCCAGCAGTACAA-  CAGTTTTGTG-3′	136
b) MLPA probesb
EXON 3	5′-GGTTAAAGGTAGGAGGCGACGAGGTGCTA-3′	5′-TCAGCGCTGAGGTCTACACGGAGGAAGAT-3′	100
IVS2_B	5′-TTCCTGGAGATCCGGTTTTGAAATGGGTCAC-3′	5′-TGTAAGGTGATCCCTCTTCTTTGGACATCAA-3′	104
IVS2_C	5′-GGGGGGTTGTGTTTCTGTTACCTTCAACTACTCCA-3′	5′-TAAGAAATGTACCAAAAACTCTTGACAGTTGGTCT-3′	112
IVS2_D	5′-GCTCTGAGTTGTCAGATTAGGTTATATTCAGATTGCT-3′	5′-GTTTTTAAAAGACTCACTAGAATCCATGAGACACTGC-3′	116
IVS2_E	5′-CGGGTCAAATGAATGAGAAAGTAGAAGATAACA-  CATTTT-3′	5′-TATTTTTTCCCTTTTACTTCAGGCACAACAACTAATTCT-3′	120
IVS6	5′-GTTCTCTAAGTATCTGTGACTGGTATTAGTGCTTAGCT-  GACCT-3′	5′-TTTTCCCTTACATCCTTACAGAGTGCTTTTTTCTTATCA-  GATA-3′	128
c) Reference genesc
SERPINB2	5′-CAGAGAACTTTACCAGCTGTGGGTTCATGCA-3′	5′-GCAGATCCAGAAGGGTAGTTATCCTGATGCG-3′	104
DACH1	5′-CTAGACCTGGAAGGCCTCCTAAGAGGACTCAAA-3′	5′-GTGTCACCTCCCCAGAGAACTCTCACATCATGCCGCA-3′	112
ING1	5′-CAGAGATCCTGAAGGAGCTAGACGAGTGCTAC-  GAGCGCT-3′	5′-TCAGTCGCGAGACAGACGGGGCGCAGAAGCGGCG-  GATGC-3′	120
SS18	5′-GACAGCATTACCAAGGACAGCAGCCACCTATGGGAAT-  GATG-3′	5′-GGTCAAGTTAACCAAGGCAATCATATGATGGGTCAGAGA-  CAGATT-3′	128

^aSequences of the 12 pairs of oligonucleotides designed to analyze the 12 exons of PRKAR1A gene.

^bSequences of the oligonucleotides designed specifically to localize the breakpoints of exons 3–6 deletion in one family of CNC.

^cSequences of the oligonucleotides designed for the 4 reference genes.