Sequential chemiluminescent detection of target DNAs without stripping and reprobing.
We present a simple method for sequential chemiluminescent detections of two different DNA loci on a single Southern blot. First, an enzyme-linked DNA probe for a unique sequence is detected with a horse-radish peroxidase (HRP) substrate followed by the detection of another enzyme-linked DNA probe for a different unique sequence with an alkaline phosphatase (AP) substrate that simultaneously inhibits the chemiluminescence generated by HRP. Such sequential detection steps eliminate the need to strip and reprobe blots and can be performed with no intervening steps.
Reddy
L V
LV
Lumigen, Inc., Southfield, MI 48034, USA.
DeSilva
R
R
Handley
R S
RS
Schaap
A P
AP
Akhavan-Tafti
H
H
D000469:Alkaline Phosphatase; D001483:Base Sequence; D001709:Biotechnology; D015139:Blotting, Southern; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004247:DNA / Q000032:analysis* / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes; D005069:Evaluation Studies as Topic; D005819:Genetic Markers; D005838:Genotype; D006735:Horseradish Peroxidase; D006801:Humans; D008163:Luminescent Measurements
C505032:CFTR protein, human; D017931:DNA Primers; D015342:DNA Probes; D005819:Genetic Markers; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA; D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase