Table 3

Assay validation on peripheral blood DNAs of NL, PM, and/or FM females and males

ID/sexSouthern (kb) or PCR (No of repeats*) resultSouthern/PCR classificationNon-Met-PCR result (bp/repeats)†Met-PCR result (bp/repeats)†mTP-PCR resultTriple ms-PCR classification
Southern blot fragments are based on EcoRI/EagI double digests hybridised with probe StB12.3, where a 30 repeat normal allele is detected as a 2.8 kb non-methylated or a 5.2 kb methylated fragment.
*Number of CGG repeats determined by PCR amplification of unmodified DNA followed by Genescan™ analysis on an ABI 310 (Applied Biosystems, Forster City).
†Rounded off to the nearest 10 bp and 5 CGG repeats.
F, female; M, male; PCR, polymerase chain reaction.
687-1/F30, 31 rpt*NL female260 bp/30 rpt200bp/30 rptNL smearNL female
FX0020/F2.8, 3.0, 5.2, 5.4 kbPM female270 and 360 bp/35 and 65 rpt200 and 300 bp/30 and 65 rptPFM smearPM female
FX0015/F2.8, 3.0, 5.2, 5.4 kbPM female270 and 410 bp/35 and 80 rpt200 and 350 bp/30 and 80 rptPFM smearPM female
FX0018/F2.8, 3.4, 5.2, 5.7 kbPM female270 and 700 bp/35 and175 rpt200 and 550 bp/30 and 145 rptPFM smearPM female
FX0017/F2.8, 5.2, 6.4 kbFM female250 bp/25 rpt190, 400 and 850 bp/25, 95 and 245 rptPFM smearFM female
FX0019/F2.8, 5.2, 6.7–8.2 kbFM female300 bp/45 rpt240, 800 and>1000 bp/45, 230 and>295 rptPFM smearFM female
FX0025/F2.8, 5.2, 7.5–8.0 kbFM female300 bp/45 rpt240 bp/45 rptPFM smearFM female
662–1/M31 rpt*NL male270 bp/35 rptno PCR productno smearNL male
FX0021/M3.1 kbPM male450 bp/95 rptno PCR productno smearPM male
FX0024/M3.3 kbPM male650 bp/160 rptno PCR productno smearPM male
FX0023/M6.2 kbFM male400 and 650 bp/75 and 160 rpt>1000 bp/>295 rptPFM smearmos FM male
194–2/M3.3, 6.3, 6.75, 6.3–8.4 kbmos FM male700 bp/175 rptno PCR productPFM smearmos FM male
FX0013/M6.6–12.0 kbFM maleno PCR product500, 600, 700 and >1000 bp/130, 165, 195 and >295 rptPFM smearFM male
FX0012/M8.8–9.1 kbFM maleno PCR productno PCR productPFM smearFM male