Improved methodology for assessment of mRNA levels in blood of patients with FMR1 related disorders.
David E Godler, Danuta Z Loesch, Richard Huggins, Lavinia Gordon, Howard R Slater, Freya Gehling, Trent Burgess, KH Andy Choo
BMC Clin Pathol | Published : 2009
BACKGROUND: Elevated levels of FMR1 mRNA in blood have been implicated in RNA toxicity associated with a number of clinical conditions. Due to the extensive inter-sample variation in the time lapse between the blood collection and RNA extraction in clinical practice, the resulting variation in mRNA quality significantly confounds mRNA analysis by real-time PCR. METHODS: Here, we developed an improved method to normalize for mRNA degradation in a sample set with large variation in rRNA quality, without sample omission. Initially, RNA samples were artificially degraded, and analyzed using capillary electrophoresis and real-time PCR standard curve method, with the aim of defining the best predi..View full abstract