A comparison of commercially-available automated and manual extraction kits for the isolation of total RNA from small tissue samplesShow full item record
Title | A comparison of commercially-available automated and manual extraction kits for the isolation of total RNA from small tissue samples |
---|---|
Author | Jeffries, Marlo K. Sellin; Kiss, Andor J.; Smith, Austin W.; Oris, James T. |
Date | 2014-11-14 |
Abstract | Background: This study compared the performance of five commercially available kits in extracting total RNA from small eukaryotic tissue samples (<15 mg). Total RNA was isolated from fathead minnow (Pimephales promelas) tissues (spleen, blood, kidney, embryo, and larvae) using the Qiagen RNeasy® Plus Mini, Qiagen RNeasy® Plus Universal, Promega Maxwell® 16 LEV simplyRNA, Ambion MagMAX™-96 and Promega SimplyRNA HT kits. Kit performance was evaluated via measures of RNA quantity (e.g., total RNA amount) and quality (e.g., ratio of absorbance at 260 and 280 nm, RNA integrity number (RIN), presence of gDNA). Results: With the exception of embryos, each kit generally extracted =5µg of total RNA from each sample. With regard to RNA quality, the RINs of RNA samples isolated via the Plus Mini and Maxwell® 16 kits were consistently higher than those of samples extracted via the remaining three kits and for all tissues, these kits produced intact RNA with average RIN values = 7. The Plus Universal and SimplyRNA HT kits produced moderately degraded (RIN values <7, but = 5), while the RNA recovered via the MagMAX™ kit tended to exhibit a high degree of degradation (RIN values <5). Conclusions: Each kit was generally capable of extracting the amount of RNA required for most downstream gene expression applications suggesting that RNA yield is unlikely to be a limiting factor for any of the kits evaluated. However, differences in the quality of RNA extracted via each of the kits indicate that these kits may differ in their ability to yield RNA acceptable for some applications. Overall, the findings of this study demonstrate that there are practical differences between commercially available RNA extraction kits that should be taken into account when selecting extraction methods to be used for isolating RNA designated for gene expression analysis. |
Link | https://doi.org/10.1186/s12896-014-0094-8
https://repository.tcu.edu/handle/116099117/11450 https://bmcbiotechnol.biomedcentral.com/articles/10.1186/s12896-014-0094-8 |
Department | Biology |
Subject | RNA extraction
Gene expression qPCR RNA-Seq Next generation sequencing Microarray Small tissues Fathead minnow |
Files in this item
This item appears in the following Collection(s)
- Research Publications [837]
Related items
Showing a few items related by title, author, creator and subject.
-
An NGQD Based Diagnostic Tool for Pancreatic Cancer
Ajgaonkar, Ryan (2021)Pancreatic cancer is extremely difficult to detect at early stages and has a low five-year-survival rate. Nanoparticles have recently emerged as frontrunners for diagnostic applications due to their small size in the 1-100nm ... -
The Effect of Zika Virus Protein NS5 on the Host Innate Immune Response
Elliott, Jacey (2020)Zika virus (ZIKV) is single stranded RNA virus that has raised global health concerns in recent years, causing diseases such as Guillain-Barré Syndrome, adult meningoencephalitis, and fetal microencephaly. ZIKV is known ... -
Itraconazole Exerts Its Antitumor Effect in Esophageal Cancer by Suppressing the HER2/AKT Signaling Pathway
Zhang W.; Bhagwath A.S.; Ramzan Z.; Williams T.A.; Subramaniyan I.; Edpuganti V.; Kallem R.R.; Dunbar K.B.; Ding P.; Gong K.; Geurkink S.A.; Beg M.S.; Kim J.; Zhang Q.; Habib A.A.; Choi S.-H.; Lapsiwala R.; Bhagwath G.; Dowell J.E.; Melton S.D.; Jie C.; Putnam W.C.; Pham T.H.; Wang D.H. (2021)Itraconazole, an FDA-approved antifungal, has antitumor activity against a variety of cancers. We sought to determine the effects of itraconazole on esophageal cancer and elucidate its mechanism of action. Itraconazole ...
© TCU Library 2015 | Contact Special Collections |
HTML Sitemap