Exon (Ex) or intron (Int) number + nucleotide position1-150 | Sequence changes (in bold; mutant underlined) | Protein change | Primers used in the first PCR1-151 | Size of product (bp) | Primers used in the second PCR1-152 | Size of product (bp) |
---|---|---|---|---|---|---|
Ex 1 | ||||||
921-150 | CCG→C T G | Pro31-Leu | P5+P48 | 1009 | P92C/P92 T +P48 (+P70) | 625 |
Int 2 | ||||||
659 | A/C→ G | Splice mutation | P5+P48 | 423 | P659A/P659 G +P48 (+P5) | 86 |
P659C/P659 G +P48 (+P5) | ||||||
Ex 4 | ||||||
1004 | ATC→A A C | Ile73-Asn | P55+P11 | 1385 | P1004T/P1004 A + P55 (+P19) | 322 |
Ex 6 | ||||||
1388 | GTG→G A G | Val238-Glu | P55+P4 | 2064 | P1388T/P1388 A +P55 (+P11) | 706 |
1394 | ATG→A A G | Met240-Glu | P55+P4 | 2064 | P1388T/P1388 A +P55 (+P11) | 706 |
1385 | ATC→A A C | Ile237-Asn | P55+P4 | 2064 | P1385T/P1385 A +P55 (+P11) | 706 |
Ex 7 | ||||||
16881-150 | GTG→ T TG | Val282-Leu | P55+P4 | 2064 | P1688G/P1688 T +P55 (+P11) | 1005 |
1718 | GGT→ A GT | Gly292-Ser | P55+P4 | 2064 | P1718G/P1718 A +P55 (+P11) | 1033 |
1768 | Ins T | Frameshift | P55+P4 | 2064 | P1768G/P1768 T +P55 (+P71) | 1078 |
Int 7 | ||||||
1784 | GT→ C T | Splice mutation | P55+P4 | 2064 | P1784G/P17884 C +P55 (+P11) | 1101 |
Ex 8 | ||||||
1999 | CAG→ T AG | Gln319-stop | P55+P4 | 2064 | P1999C/P1999 T +P55 (+P11) | 1317 |
2113 | CGG→ T GG | Arg357-Trp | P55+P4 | 2064 | P2113C/P2113 T +P55 (+P11) | 1430 |
Ex 9 | ||||||
2344 | TGG→T A G | Trp406-stop | P55+P4 | 2064 | P2344G/P2344 A +P55 (+P11) | 1660 |
Ex 10 | ||||||
25841-150 | CCC→ T CC | Pro454-Ser | P45+P4 | 619 | P2584C/P2584 T +P45 (+P11) | 455 |
2675 | CGG→C C G | Arg484-Pro | P47+P4 | 237 | P2675G/P2675 C +P47 (+P11) | 165 |
2675–2676 | CGG→C C | Arg484-frameshift | P47+P4 | 237 | P2675G/P2676 del +P47 (+P71) | 165 |
Primer names are as described by Wedell and Luthman.33
↵1-150 Mutations associated with non-classical 21-hydroxylase deficiency.
↵1-151 P48 or P55 were used as CYP21 specific primers.
↵1-152 Allele specific primers are indicated in pairs with the normal sequence represented by bold lettering at the end of the primer name while the mutant sequence specific primer ends with bold and underlined lettering. Control primers are indicated in brackets (see Methods).