Detection of sub-picogram quantities of specific DNA sequences on blot hybridization with biotinylated probes.
A sensitive method for detecting biotinylated DNA probes on dot and Southern blots is described which is based on the principle outlined by Leary et al (1). This system has two main components: detection of biotinylated DNA by a two-step procedure with streptavidin and poly(alkaline phosphatase); and blocking background with Tween 20. 32fg and 80fg of lambda phage DNA was detected on dot and Southern blot hybridizations respectively. 150fg of beta-globin was detected on Southern blots of genomic DNA. This method is fast, reproducible and can detect single copy genes in 0.25 micrograms genomic DNA on Southern blots.
Chan
V T
VT
Fleming
K A
KA
McGee
J O
JO
D000469:Alkaline Phosphatase; D001426:Bacterial Proteins; D010582:Bacteriophage lambda / Q000032:analysis; D001710:Biotin; D004247:DNA / Q000032:analysis*; D004279:DNA, Viral / Q000032:analysis; D005914:Globins / Q000032:analysis; D009693:Nucleic Acid Hybridization; D011136:Polysorbates; D019809:Streptavidin
D001426:Bacterial Proteins; D004279:DNA, Viral; D011136:Polysorbates; D001710:Biotin; D005914:Globins; D004247:DNA; D019809:Streptavidin; D000469:Alkaline Phosphatase