کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
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3407067 | 1593495 | 2010 | 5 صفحه PDF | دانلود رایگان |

Protecting RNA from degradation, whilst maintaining its biological activity, is essential in molecular biology. However, RNA is very sensitive to degradation by ribonucleases, especially at temperatures above 0 °C. The stability of RNA was examined at 4 °C and −20 °C, in a new stabilizing solution consisting of a low-molarity mixture of chaotropic agents guanidinium and ammonium thiocyanate, a buffer for pH stabilization, phenol, and yeast RNA. Two substrates were tested for storage: RNA in human plasma positive for hepatitis C virus (HCV) and naked RNA (purified from HCV positive human plasma or transcribed in vitro). Stability was followed by viral load estimation, using an in-house competitive RT-PCR assay. Naked RNA purified from human plasma positive for HCV was stable at 4 °C for at least 24 months. An RNA standard transcribed in vitro was still viable after 36 months of storage at 4 °C. Human plasma dilutions positive for HCV were stable for at least 5 months in this solution when stored at 4 °C. It was concluded that the described stabilizing solution ensures long-term stability on naked RNA at 4 °C, and ideal for the storage of RNA controls and standards for molecular diagnosis, the solution may be used for preserving clinical samples prior to transport to a clinical laboratory.
Journal: Journal of Virological Methods - Volume 168, Issues 1–2, September 2010, Pages 207–211