A rapid and convenient method to prepare DIG-labelled RNA probes for use in non-radioactive in situ hybridization.
We describe here the use of PCR-generated templates incorporating T3 polymerase sites in order to prepare digoxigenin (DIG)-labelled cRNA probes against any gene of known sequence. This method was applied to the preparation of probes specific for chicken glyceraldehyde-3-phosphate dehydrogenase messenger RNAs and we demonstrate that such probes can be used for in situ hybridization (ISH). This technique therefore represents a rapid and convenient means to prepare DIG-labelled cRNA probes for use in a non-radioactive ISH. It adds speed and convenience of probe preparation to the previously described advantages of non-radioactive detection techniques.
Gandrillon
O
O
Département de Biologie Moléculaire et Cellulaire, Ecole Normale Supérieure de Lyon, France.
Solari
F
F
Legrand
C
C
Jurdic
P
P
Samarut
J
J
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D004076:Digoxigenin; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D017403:In Situ Hybridization / Q000379:methods*; D008264:Macrophages; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D018075:RNA, Complementary; D012333:RNA, Messenger / Q000032:analysis
D018075:RNA, Complementary; D012333:RNA, Messenger; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D004076:Digoxigenin