Determination of half-life for mRNA expressed in few copies and with short life.
Determination of stability of mRNA expressed in low abundance and with short life requires a sensitive method for detection and an accurate measurement of mRNA level. The method described here meets both of these criteria. An uniformly labeled single-stranded probe with high specific activity was used for the increased sensitivity in detection and quantitation of mRNA. The levels of mRNA were normalized with an invariant mRNA specie with longer half-life under the same experimental conditions in the same cells. These modifications may be helpful for situations where measurement of mRNA half-life for short-lived few copies of mRNA is necessary.
Raval
P
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Department of Pharmacology, University of Nebraska Medical Center, Omaha, 68198-6260, USA.
D000818:Animals; D001483:Base Sequence; D001853:Bone Marrow / Q000187:drug effects / Q000378:metabolism; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D046508:Culture Techniques; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes; D004277:DNA, Single-Stranded / Q000737:chemistry; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D006207:Half-Life; D016899:Interferon-beta / Q000235:genetics* / Q000378:metabolism; D007553:Isotope Labeling; D008070:Lipopolysaccharides / Q000633:toxicity; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*
D017931:DNA Primers; D015342:DNA Probes; D004277:DNA, Single-Stranded; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D016899:Interferon-beta; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases