A Method of Low-Temperature Storing of Agarose Slides with Lysed Cells

被引:0
|
作者
Kuznetsova, E. A. [1 ]
Dyukina, A. R. [1 ]
Chernigina, I. A. [1 ]
Sirota, N. P. [1 ]
机构
[1] Russian Acad Sci, Inst Theoret & Expt Biophys, Pushchino 142292, Moscow Region, Russia
关键词
leukocyte DNA damage; storing slides; comet assay; ALKALINE COMET ASSAY; DNA-DAMAGE;
D O I
10.1007/s10517-013-2245-7
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
A method has been developed for a long-term low-temperature storage (-10 to -15A degrees C) of the agarose slides with nucleoids (lysed eukaryotic cells). After lysis of agarose-immobilized cells, the slides were incubated for 30 min in phosphate buffer with 50% glycerol and 100 mM EDTA, thereupon they were stored in a freezer at -10 to -15A degrees C. After long-term storage, the slides were re-incubated for 30 min in lysing solution. The measurements of the baseline and in vitro induced DNA damage in nucleoids of the human and mouse leukocytes, which had been stored in agarose slides at low temperature, showed that DNA damage level determined after a 40-day storage did not significantly differ from that of the fresh slides. The advanced storage method is simple and reliable; it opens the way to avoid cryopreservation of the biological samples and to process little by little a great number of the identically prepared slides.
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页码:757 / 759
页数:3
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