PMC

A Southern blot analysis of rolling circle amplification (RCA) products, probed with DIG-labeled (CAG)₇ LNA probe (left), or ³²P-labeled DMPK probe (right). Lane 1, DM1 fibroblast cells containing 900 repeats; lane 2, DM1 cerebellum; lanes 3 and 4, DM1 leukocytes from two patients; lanes 5,6, and 7, leukocytes from three normal controls. Arrows indicate normal alleles (807 bp plus repeats). M, DIG-labeled molecular weight marker.
B Southern blot of RCA products from serially diluted template DNA of DM1 cerebellum. The products were probed with DIG-labeled (CAG) ₇ LNA probe. Expanded repeats are observed in the RCA products from 20 ng of template DNA.

A Restriction map of the genomic locus encompassing the CCTG repeat in ZNF9.
B Southern blot of leukocyte DNA from DM2 patients and controls using DIG-labeled (CCTG)₅ LNA probe. Five hundred ng (left) or three µg (right) of genomic DNA digested with AluI and HaeIII were loaded in each well. Lanes 1–4 are from patients with DM2, lanes 5–7 are from healthy controls. Asterisk indicates nonspecific signal, M indicates DIG-labeled molecular weight marker.
C Southern blot showing serial dilution of DM2 leukocyte DNA hybridized with DIG-labeled (CCTG) ₅ LNA probe. The repeat expansion is detected in 50 ng of genomic DNA.
D Repeat length comparison between muscle and leucocyte DNA from two patients with DM2. The repeat size was considerably larger in muscle than blood for both patients.

A Restriction map of the DM1 locus. For each restriction enzyme, the cleavage site closest to the CTG repeat is indicated.
B Southern blot analysis of genomic DNA digested by BglI. Left panel shows DNA probed with digoxigenin (DIG)-labeled (CAG)₇ locked nucleic acid (LNA) probe. Lane 1, DM1 skeletal muscle (300 ng of DNA); lane 2, DM1 heart (300 ng); lane 3, DM1 cerebellum (200 ng); lane 4, DM1 fibroblast cells containing 2000 repeats (300 ng); lane 5, congenital DM1 (CDM) heart (300 ng); lanes 6 and 7, normal leukocytes (300 ng); M indicates DIG-labeled molecular weight marker. Right panel shows the same DNA samples analyzed with ³²P-labeled p5B1.4 probe. Lane 1, DM1 skeletal muscle (4 µg of DNA); lane 2, DM1 heart (4 µg); lane 3, DM1 cerebellum (3 µg); lane 4, DM1 fibroblast cells containing 2000 repeats (4 µg); lane 5, CDM heart (4 µg); lanes 6 and 7, normal leukocytes (6 µg). The arrow indicates the normal DMPK allele (3.3 kb).
C Southern blot showing serial dilution of genomic DNA obtained from congenital DM1 heart, hybridized with DIG-labeled (CAG) ₇ LNA probe. Genomic DNA was digested with HaeIII. The repeat expansion is clearly detected in 50 ng of genomic DNA.
D Southern blot of cardiac DNA digested with different restriction enzymes. Expanded alleles are almost same size after digestion with each of three different 4-base cutters (DpnII, AluI, and HaeIII). Signals were not detected after digest with MwoI, a restriction enzyme that cuts within the CTG tract. No signal was seen in control samples.