BACKGROUND Whole blood expression profiling is frequently performed using PAXgene (Qiagen) or Tempus (Life Technologies) tubes. Here, we compare 6 novel generation RNA isolation protocols with respect to RNA quantity, quality and recovery of mRNA and miRNA. METHODS 3 PAXgene and 3 Tempus Tubes were collected from participants of the LIFE study with (n = 12) and without (n = 35) acute myocardi...

The article refers to the technique of biomagnetical separation (BS) of mRNA using "Dynabeads mRNA DIRECT". After a brief survey of RNA isolation methods authors describe a general procedure of BS as well as its concrete application for mRNA isolation from bronchoalveolar cells. Authors report results of specific experiments to assess the efficiency and accuracy of mRNA isolation by BS. The app...

.......................................................................................................36 Introduction..................................................................................................37 Materials and Methods.................................................................................38 Animal Management...........................................................

RNA isolation is difficult in plants that contain large amounts of polysaccharides and polyphenol compounds. To date, no commercial kit has been developed for the isolation of high-quality RNA from tissues with these characteristics, especially for fruit. The common protocols for RNA isolation are tedious and usually result in poor yields when applied to recalcitrant plant tissues. Here an effi...

Reverse transcription-PCR (RT-PCR) has traditionally required time-consuming RNA extraction and purification. This report demonstrates that one can completely avoid the RNA extraction step in RT-PCR by basing the comparison of samples on cell number rather than micrograms of total RNA. A new method for lysing cells while preserving RNA is described. RT-PCR is carried out (i) by rapidly freezing...

Conventional RNA isolation methods optimised for pine seedlings have been shown to produce poor quality RNA when applied to needles of adult pine trees. We describe here a modified procedure to isolate high pure RNA from needles of thirty-year-old maritime pines, exhibiting high levels of phenolics, polysaccharides and RNases. Major changes were the inclusion of proteinase K in the extraction m...

This chapter provides detailed methodologies for isolating total RNA and polyadenylated RNA from E. coli and other prokaryotes, along with the procedures necessary to analyze the processing and decay of specific transcripts and determine their 3'- and 5'-ends. The RNA isolation methods described here facilitate isolating good-quality RNA in a very cost-effective way compared to the commercially...

MicroRNAs (miRNAs) are present in all known plant and animal tissues and appear to be somewhat concentrated in the mammalian nervous system. Many different miRNA expression profiling platforms have been described. However, relatively little research has been published to establish the importance of 'upstream' variables in RNA isolation for neural miRNA expression profiling. We tested whether ap...