OBJECTIVE The preservation of biological samples at a low temperature is important for later biochemical and/or histological analyses. However, the molecular viability of thawed samples has not been studied sufficiently in depth. The present study was undertaken to evaluate the viability of intact tissues, tissue homogenates, and isolated total RNA after defrosting for more than twenty-four hou...

The small subunit (SSU) ribosomal RNA (rRNA) is the most commonly used marker for identification of microbial taxa, but its full-length reconstruction from high-throughput sequencing (HTS) data remains challenging. Metagenomics and total (total RNA-Seq) are target-PCR-free HTS methods that to characterize communities simultaneously reconstruct SSU rRNA sequences. However, more testing required ...

Abstract Background The reproducibility of gene expression measured by RNA sequencing (RNA-Seq) is dependent on the depth. While unmapped or non-exonic reads do not contribute to quantification, duplicate quantification but are informative for reproducibility. We show that mapped, exonic, non-duplicate (MEND) a useful measure RNA-Seq datasets used analysis. Findings In bulk from 2,179 tumors in...

This report describes a newly developed method using Trizol LS reagent that can reliably extract high quality total RNA from frozen human leukaemic bone marrow samples. Extraction of total RNA from 71 frozen leukaemic bone marrow samples obtained at the time of diagnosis produced a median yield of 145 micro g/ml leukaemic bone marrow. Total RNA samples could be reverse transcribed into cDNA and...

Isolating RNA from insects is becoming increasingly important in molecular entomology. Four methods including three commercial kits RNeasy Mini Kit (Qiagen), SV Total RNA isolation system (Promega), TRIzol reagent (Invitrogen), and a cetyl trimethylammonium bromide (CTAB)-based method were compared regarding their ability to isolate RNA from whole-body larvae of Thaumatotibia leucotreta (Meyric...

The assessment of RNA integrity is a critical first step in obtaining meaningful gene expression data. Working with low-quality RNA may strongly compromise the experimental results of downstream applications which are often labour-intensive, time-consuming, and highly expensive. Using intact RNA is a key element for the successful application of modern molecular biological methods, like qRT-PCR...

The amount, by mass, of poly(A+) mRNA present in the polyribosomes of third-instar larvae of Drosophila melanogaster, and the relative contribution of the poly(A+) mRNA to the sequence complexity of total polysomal RNA, has been determined. Selective removal of poly(A+) mRNA from total polysomal RNA by use of either oligo-dT-cellulose, or poly(U)-sepharose affinity chromatography, revealed that...