The Roslin Institute, University of Edinburgh, Edinburgh, UK.
Neuropathol Appl Neurobiol. 2011 Oct;37(6):633-42. doi: 10.1111/j.1365-2990.2011.01162.x.
To determine premortem and post mortem factors affecting quality and yield of RNA isolated from the unique archived brain material in the UK National Creutzfeldt-Jakob Disease Surveillance Unit Brain and Tissue Bank and to compare this to control brain tissue with no neurological disease.
In parallel and in replicate, RNA was prepared from the frontal parasagittal or subfrontal cortex of samples dissected from half brains (frozen intact) or from brain samples snap frozen or placed in RNALater. A total of 350 RNA samples from 78 human autopsy cases, 21 variant Creutzfeldt-Jakob disease, 26 other neurological diseases and 31 non-neurological diseases were studied.
There was no difference in the quality or yield of RNA isolated from variant Creutzfeldt-Jakob disease, other neurological disease and non-neurological disease brains. RNA preparations from archived frozen half brains or snap frozen autopsy samples were generally of poor quality (RNA integrity number<5). There was a highly significant negative correlation between the number of times frozen half brains had been sampled and the quality of RNA. Samples stored in RNALater provided higher-quality RNA (RNA integrity number>5). Age at death, gender, post mortem interval and freezer storage time had no effect on RNA quality.
Reasonable-quality RNA can be isolated from samples dissected from archived frozen human half brains but repeated sampling results in RNA degradation. Better-quality RNA is obtained from samples placed in RNALater than from snap frozen samples. The quality and yield of RNA are not affected by age at death, gender, post mortem interval of >6 h or freezer storage time.
确定影响从英国国家克雅氏病监测单位脑和组织库中独特存档脑组织中分离的 RNA 质量和产量的生前和死后因素，并将其与无神经疾病的对照脑组织进行比较。
平行且重复地，从对半脑（完整冷冻）进行解剖或对快速冷冻或置于 RNALater 中的脑样本中，从额叶旁矢状或额下皮质制备 RNA。共研究了 78 例尸检病例、21 例变异型克雅氏病、26 例其他神经疾病和 31 例非神经疾病的 350 个 RNA 样本。
从变异型克雅氏病、其他神经疾病和非神经疾病脑分离的 RNA 的质量或产量没有差异。来自存档冷冻半脑或快速冷冻尸检样本的 RNA 制剂通常质量较差（RNA 完整性编号<5）。冷冻半脑被取样的次数与 RNA 质量之间存在显著的负相关。储存在 RNALater 中的样本提供了更高质量的 RNA（RNA 完整性编号>5）。死亡年龄、性别、尸检间隔时间和冷冻储存时间对 RNA 质量没有影响。
可以从存档冷冻的人类半脑解剖样本中分离出合理质量的 RNA，但重复取样会导致 RNA 降解。与快速冷冻样本相比，放置在 RNALater 中的样本获得的 RNA 质量更好。RNA 的质量和产量不受死亡年龄、性别、>6 小时的尸检间隔时间或冷冻储存时间的影响。
