The effect of long-term -80°C storage of thyroid biospecimens on RNA quality and ensuring fitness for purpose.
The effect of long-term -80°C storage of thyroid biospecimens on RNA quality and ensuring fitness for purpose.
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DOI：
10.1136/jclinpath-2016-203697
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发表时间：
2016-12
影响因子：
3.4
通讯作者：
Thomas GA
中科院分区：
文献类型：
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作者：
Mathieson W;Betsou F;Myshunina T;Pushkarev V;Pushkarev V;Shinkarkina A;Voskoboynyk L;Thomas GA
为了确定核糖核酸在−80°C的长期储存中是否会降解，以及核糖核酸完整性数(RIN)是否决定了严重降解的核糖核酸的“适用性”。549例甲状腺标本在−80℃下保存0.1~10.9年，其RIN与保存时间呈正相关。用RT-PCR方法对65、265、534和942个碱基对的羟甲基丁烷合成酶的扩增产物长度进行了测定，并测定了冷冻保存和RIN等降解的FFPE生物样品的RNA长度。贮藏时间与RIN无关。与相同RIN的FFPE样品相比，冷冻保存的样品可获得更长的扩增片段。在−80°C下储存较长时间的甲状腺生物样品中，核糖核酸不会降解。虽然已知RIN可以预测高质量样品对分析平台的适应性，但这一预测在严重降解的样品中是不可靠的。
To establish whether RNA degrades in long-term storage at −80°C and whether RNA integrity numbers (RINs) determine ‘fitness for purpose’ in severely degraded RNA. RNA was extracted from 549 thyroid biospecimens stored at −80°C for 0.1–10.9 years then their RINs correlated with storage time. RT-PCR for 65, 265, 534 and 942 base pair amplicons of hydroxymethylbilane synthase was used to measure amplicon length in RNA from cryopreserved and FFPE biospecimens that were equally degraded according to RIN. Storage time did not correlate with RIN. Longer amplicons were obtained from cryopreserved samples than FFPE samples with equal RINs. RNA does not degrade in thyroid biospecimens stored for long periods of time at −80°C. Although RINs are known to predict amenability to analytical platforms in good quality samples, this prediction is unreliable in severely degraded samples.
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影响因子：
--
作者：
Schroeder A;Mueller O;Stocker S;Salowsky R;Leiber M;Gassmann M;Lightfoot S;Menzel W;Granzow M;Ragg T
通讯作者：
Ragg T
影响因子：
3.7
作者：
Rudloff U;Bhanot U;Gerald W;Klimstra DS;Jarnagin WR;Brennan MF;Allen PJ
通讯作者：
Allen PJ
影响因子：
2.7
作者：
Opitz L;Salinas-Riester G;Grade M;Jung K;Jo P;Emons G;Ghadimi BM;Beissbarth T;Gaedcke J
通讯作者：
Gaedcke J
影响因子：
1.6
作者：
Andreasson, Adam;Kiss, Nimrod B.;Hoog, Anders
通讯作者：
Hoog, Anders
影响因子：
10.4
作者：
Hebels DG;Georgiadis P;Keun HC;Athersuch TJ;Vineis P;Vermeulen R;Portengen L;Bergdahl IA;Hallmans G;Palli D;Bendinelli B;Krogh V;Tumino R;Sacerdote C;Panico S;Kleinjans JC;de Kok TM;Smith MT;Kyrtopoulos SA;EnviroGenomarkers Project Consortium
通讯作者：
EnviroGenomarkers Project Consortium