A simple method for quantifying specific mRNAs in small numbers of early mouse embryos.
Amplification of sequences by the polymerase chain reaction (PCR) has become a powerful tool in the study of gene expression. The technique is, in fact, so powerful that it may detect 'leaky transcription'. Thus, it is now important to be able to quantify the transcripts that are amplified to determine whether or not they represent legitimate transcription of target genes. In this paper, we describe a one-step amplification reaction coupled to solution hybridization/RNase protection that is capable of quantitating specific transcripts in total RNA from one to ten preimplantation mouse embryos and is generally applicable to any cloned mRNA sequence.
Gaudette
M F
MF
Cell Biology Group, Worcester Foundation for Experimental Biology, Shrewsbury, MA 01545.
Crain
W R
WR
D000818:Animals; D016375:Antisense Elements (Genetics); D001483:Base Sequence; D001755:Blastocyst / Q000378:metabolism*; D003001:Cloning, Molecular; D004247:DNA; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis*; D006163:Ribonuclease T1; D014158:Transcription, Genetic
D016375:Antisense Elements (Genetics); D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D004247:DNA; D006163:Ribonuclease T1