High DNA stability in white blood cells and buffy coat lysates stored at ambient temperature under anoxic and anhydrous atmosphere

被引:9
作者
Fabre, Anne-Lise [1 ,2 ]
Luis, Aurelie [1 ]
Colotte, Marthe [2 ]
Tuffet, Sophie [1 ]
Bonnet, Jacques [1 ,3 ]
机构
[1] Univ Bordeaux, R&D Dept, Imagene, ENSTBB, 146 Rue Leo Saignat, Bordeaux, France
[2] Prod Platform, Imagene, Rue Henri Desbrueres,Genopole Campus 1,Bat 6, Evry, France
[3] Univ Bordeaux, INSERM, U1218, Inst Bergonie, 229 Cours Argonne, Bordeaux, France
关键词
LYOPHILIZED LIPID/DNA COMPLEXES; PERIPHERAL-BLOOD; QUALITY-CONTROL; NUCLEIC-ACIDS; GENOMIC DNA; STORAGE; DEGRADATION; BIOSPECIMENS; AMPLIFICATION; PRESERVATION;
D O I
10.1371/journal.pone.0188547
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Conventional storage of blood-derived fractions relies on cold. However, lately, ambient temperature preservation has been evaluated by several independent institutions that see economic and logistic advantages in getting rid of the cold chain. Here we validated a novel procedure for ambient temperature preservation of DNA in white blood cell and buffy coat lysates based on the confinement of the desiccated biospecimens under anoxic and anhydrous atmosphere in original hermetic minicapsules. For this validation we stored encapsulated samples either at ambient temperature or at several elevated temperatures to accelerate aging. We found that DNA extracted from stored samples was of good quality with a yield of extraction as expected. Degradation rates were estimated from the average fragment size of denatured DNA run on agarose gels and from qPCR reactions. At ambient temperature, these rates were too low to be measured but the degradation rate dependence on temperature followed Arrhenius' law, making it possible to extrapolate degradation rates at 25 degrees C. According to these values, the DNA stored in the encapsulated blood products would remain larger than 20 kb after one century at ambient temperature. At last, qPCR experiments demonstrated the compatibility of extracted DNA with routine DNA downstream analyses. Altogether, these results showed that this novel storage method provides an adequate environment for ambient temperature long term storage of high molecular weight DNA in dehydrated lysates of white blood cells and buffy coats.
引用
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页数:23
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