Yield and Integrity of RNA from Brain Samples are Largely Unaffected by Pre-analytical Procedures

被引:3
|
作者
Jensen, Pernille Sos Hovgaard [1 ]
Johansen, Maja [1 ]
Bak, Lasse K. [2 ]
Jensen, Lars Juhl [3 ]
Kjaer, Christina [1 ,2 ]
机构
[1] Univ Coll Copenhagen, Fac Hlth & Technol, Dept Technol, DK-2200 Copenhagen, Denmark
[2] Univ Copenhagen, Fac Hlth & Med Sci, Dept Drug Design & Pharmacol, DK-2100 Copenhagen, Denmark
[3] Univ Copenhagen, Fac Hlth & Med Sci, Novo Nordisk Fdn Ctr Prot Res, Dis Syst Biol Program, DK-2200 Copenhagen, Denmark
关键词
RNA isolation; RNA Integrity Number (RIN); Brain tissue; Pre-analytical factors; Gene expression; RNA stability; RNA degradation; GENE-EXPRESSION; TISSUE; QUALITY; PRESERVATION; STABILITY; STABILIZATION; MORPHOLOGY; FROZEN;
D O I
10.1007/s11064-020-03183-z
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Gene expression studies are reported to be influenced by pre-analytical factors that can compromise RNA yield and integrity, which in turn may confound the experimental findings. Here we investigate the impact of four pre-analytical factors on brain-derived RNA: time-before-collection, tissue specimen size, tissue collection method, and RNA isolation method. We report no significant differences in RNA yield or integrity between 20 mg and 60 mg tissue samples collected in either liquid nitrogen or the RNAlater stabilizing solution. Isolation of RNA employing the TRIzol reagent resulted in a higher yield compared to isolation via the QIAcube kit while the latter resulted in RNA of slightly better integrity. Keeping brain tissue samples at room temperature for up to 160 min prior to collection and isolation of RNA resulted in no significant difference in yield or integrity. Our findings have significant practical and financial consequences for clinical genomic departments and other laboratory settings performing large-scale routine RNA expression analysis of brain samples.
引用
收藏
页码:447 / 454
页数:8
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