Poláková H, Kádasi L, Zelinková M
Bratisl Lek Listy. 1989 Nov;90(11):844-7.
The effect of different conditions of blood sample storage on the yield and quality of the extracted DNA was studied. Samples of whole umbilical blood were divided into eight groups containing 10 samples each and stored as follows: A--no storage, DNA extracted immediately; B--24 hours at 4 degrees C; C--24 hours at room temperature; D--7 days at 4 degrees C; E--3 months at -18 degrees C; F--6 months at -70 degrees C; G--14 days at -70 degrees C, the samples were thawed one time and frozen again; H--14 days at -70 degrees C, the samples were 3 times thawed and frozen. DNA was isolated and yields quantified by spectrophotometry at 260 and 280 nm. The average DNA yield in the individual groups ranged from 574.5 to 1,075 micrograms per 10 ml of whole blood. Although variations in the yield of DNA were observed both within and among groups, there were no significant changes with respect to different storage conditions. However group H yielded significantly more DNA (900 micrograms/10 ml, p less than 0.05) compared to the control group A. After digestion with restriction endonuclease and electrophoresis in agarose gel, all DNA preparations were found to be of high molecular weight and in digestible condition. The results can be used to advantage in storing DNA for purposes of molecular diagnosis of genetic diseases.
研究了血样不同保存条件对提取DNA产量和质量的影响。将全脐血样本分成八组，每组10份样本，保存方式如下：A组——不保存，立即提取DNA；B组——4℃保存24小时；C组——室温保存24小时；D组——4℃保存7天；E组——-18℃保存3个月；F组——-70℃保存6个月；G组——-70℃保存14天，样本解冻一次后再冷冻；H组——-70℃保存14天，样本解冻3次后再冷冻。采用分光光度法于260和280nm波长处分离DNA并对产量进行定量。各小组中每10ml全血的平均DNA产量在574.5至1075微克之间。尽管组内和组间均观察到DNA产量存在差异，但不同保存条件下并无显著变化。然而，与对照组A相比，H组产生的DNA显著更多（900微克/10ml，p<0.05）。经限制性内切酶消化并在琼脂糖凝胶中电泳后，发现所有DNA制品均为高分子量且可消化状态。这些结果可有效地用于为遗传病分子诊断而保存DNA。
