Highly informative single-stranded conformation polymorphism (SSCP) of short tandem repeats in DNA identification.
The analysis of short tandem repeats (STR) by PCR is a useful technique widely used in DNA identification. Some loci have alleles differing in the number of complete repeat units, whereas others are polymorphic with alleles differing even to the level of a single base. Minor differences in the sequence of STR can confuse the analysis when the PCR product are poorly resolved in denaturing PAGE. The study by SSCP of STR in loci CSF1PO, TPOX, and THO1 and the alleles of locus DIS80 is shown in a representative case of paternity dispute. The analysis of short tandem repeats (STR) by the conformation polymorphisms of single-stranded DNA (SSCP) clearly enhanced the degree of resolution. The high resolution of SSCP of the STR may be the result of the arrangement of very prominent secondary structure and conformation due to the primary repetitive sequence. The method may be of particular interest to distinguish STR with similar PAGE mobility under denaturing conditions and to distinguish tandem repeats of the same size but different sequence.
Lucas
M
M
Departmento de Bioquímica Médica y Biología Molecular, Laboratorio de Genética Molecular, Sevilla, Spain.
Muñoz
C
C
Pintado
E
E
Solano
F
F
D000483:Alleles; D004277:DNA, Single-Stranded / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005554:Forensic Medicine; D006239:Haplotypes; D006801:Humans; D008297:Male; D010334:Paternity; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics*
D004277:DNA, Single-Stranded