Small RNA extractionSmall RNA quantificationIn spite of the growing interest in the roles and applications of small RNAs (sRNAs), sRNA isolation methods are inconsistent, tedious, and dependent on the starting number of cells. In this work, we employ ITP to isolate sRNAs from the cell鈥恖ys...
Northern blots to detect (A) RssR and (B) STnc1740 transcription. Left side: RNA was isolated from cells grown to OD600 = 0.3 in MM with 27.8 mM glucose or with 55.5 mM MI. Right side: RNA isolated from strain 14028 and its deletion mutants lackinghfq,ssrB, STnc1740,iol...
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et al. Small RNA sequencing across diverse biofluids identifies optimal methods for exRNA isolation. Cell 177, 446–462 (2019). Article CAS PubMed PubMed Central Google Scholar Thery, C., Amigorena, S., Raposo, G. & Clayton, A. Isolation and characterization of exosomes from cell culture ...
M1 Macrophages were defined as F4/80+CD11b+CD86+ cells, and M2 Macrophages were defined as F4/80+ CD11b+ CD206+ cells. 2.7. RNA extraction and quantitative reverse transcription-PCR Total RNA was extracted by Tripure Isolation Reagent (Roche, Basel Switzerland). For evaluation of mRNA ...
The 4.5S RNA of E. coli and its equivalent in B. subtilis, called small cytoplasmic RNA (scRNA), are part of the signal recognition particle (SRP) whose function is to direct proteins bearing an N-terminal export sequence to the plasma membrane as they emerge from the ribosome. This RN...
The isolation of spermatozoal RNA is a challenging procedure due to the intrinsic heterogeneous population of cells present in the ejaculate and the small quantity of RNA present in sperm. The transcriptome of these gametes includes a wide variety of messenger RNAs (mRNAs), small noncoding RNAs ...
It also permitted smooth integration with on-chip RT-PCR, which simultaneously detected the target microRNA,mir-191, expressed in fewer than 10 cancer cells. Our results have demonstrated that the M&M chip device is a valuable and cost-saving platform for studying small RNA expression patterns ...
We further show, using unique molecular identifiers, that stochastic capture of low input RNA rather than PCR amplification influences the biased quantitation of intermediately and lowly expressed microRNAs. Our improved method allows the processing of tens to hundreds of samples simultaneously while ...
RNA was extracted from cells using the mirVana miRNA isolation kit (Ambion) following the manufacturer's instructions. Small RNA libraries for NGS were prepared using the small RNA preparation protocol of the SOLiD Total RNA-Seq kit as per the manufacturer's instructions. Specifically, size ...