Use of biotinylated probes for detecting sickle cell anemia.
Earlier, we reported that the 5' end of the normal beta-globin gene (beta) resides on a 1.14-kilobase DNA fragment, whereas the 5' end of the sickle cell gene (beta s) resides on a 1.34-kilobase fragment. In that blot hybridization analysis, we used genomic DNA digested with restricted endonuclease Mst II, and radioactive probes with short half-life. We demonstrate here that, if a biotinylated probe is used instead in a slightly modified procedure, sickle cell anemia can be quickly and directly detected if there is as much as 5 micrograms of total genomic DNA in the sample. This procedure obviates the special precautions necessary when radioactive materials are used.
Garbutt
G J
GJ
Wilson
J T
JT
Schuster
G S
GS
Leary
J J
JJ
Ward
D C
DC
D000755:Anemia, Sickle Cell / Q000175:diagnosis* / Q000235:genetics; D001710:Biotin / Q000031:analogs & derivatives*; D003124:Colorimetry; D004247:DNA / Q000032:analysis; D004262:DNA Restriction Enzymes / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific; D003856:Deoxyuracil Nucleotides; D004587:Electrophoresis, Agar Gel; D005914:Globins / Q000235:genetics*; D006801:Humans; D008722:Methods; D009693:Nucleic Acid Hybridization
D003856:Deoxyuracil Nucleotides; D001710:Biotin; C045931:biotin-11-dUTP; D005914:Globins; D004247:DNA; D004262:DNA Restriction Enzymes; C034726:endodeoxyribonuclease MstII; D015252:Deoxyribonucleases, Type II Site-Specific