Dideoxy fingerprinting (ddE): a rapid and efficient screen for the presence of mutations.
We describe dideoxy fingerprinting (ddF), a hybrid between dideoxy sequencing and SSCP that can detect the presence of single base and other sequence changes in PCR-amplified segments. As implemented herein, ddF involves a Sanger sequencing reaction with one dideoxynucleotide followed by nondenaturing gel electrophoresis. When ddF was used to examine segments of the human factor IX gene, 84 of 84 different mutations were detected with a very low rate of false positive signals. The approximate locations of the sequence changes could be determined from the ddF pattern and samples with different sequence changes had different fingerprints. In addition, large segments could be amplified and rapidly screened by ddF in multiple smaller subsegments. The patterns observed with ddF are instructive in that they suggest an inherent limitation in the detection of certain mutations by SSCP.
Sarkar
G
G
Department of Biochemistry and Molecular Biology, Mayo Clinic/Foundation, Rochester, Minnesota 55905.
Yoon
H S
HS
Sommer
S S
SS
D016172:DNA Fingerprinting / Q000379:methods*; D004252:DNA Mutational Analysis; D004277:DNA, Single-Stranded / Q000032:analysis; D005164:Factor IX / Q000235:genetics; D006801:Humans; D009690:Nucleic Acid Conformation; D016133:Polymerase Chain Reaction / Q000379:methods*; D016037:Single-Blind Method
D004277:DNA, Single-Stranded; D005164:Factor IX