A comparative analysis of kits for nucleic acid extraction from animal blood and tissue samples

Aim. To conduct a comparative analysis of the effectiveness of com­mer­cial kits for deoxyribonucleic acid (DNA) extraction from blood and tissue, as well as ribonucleic acid (RNA) from mouse tissue, and to eva­luate the effect of the postmortem interval (PMI) on the quality and quan­tity of extracted nucleic acids (NA).

Material and methods. Blood and tissue samples (brain, heart, liver, kid­neys, lungs, muscle, and testes) from 12 Balb/C mice were used. Samp­les were collected immediately (0 h) and 24 h after euthanasia. Ten commercial kits (Russian and international) were tested for DNA ex­trac­tion, and the RNeasy Mini Kit (Qiagen, Germany) was used for RNA extraction. Quality control included measuring the concen­tra­tion (fluorometry), purity (spectrophotometry), and integrity (electro­pho­resis) of the DNA.

Results. Spin-column-­based kits (QIAamp DNA Blood Mini Kit, Qiagen, USA, D-Blood-5) demonstrated the highest DNA yield from blood. For tissue, magnetic particle kits (GM Tissue, Tissue M) provided the highest DNA yield, but the column-­based method (QIAamp DNA Mini Kit, Qiagen, Germany) yielded better purity and stability with increasing PMI. Increasing the PMI to 24 hours resulted in a significant decrease in the concentration and integrity of both DNA and RNA in all tissues except the testes. The most pronounced RNA degradation (a decrease in RNA integrity number to 2,5-2,8) was observed in the kidneys and muscles.

Conclusion. The efficiency of NA isolation significantly depends on the type of kit, biomaterial, and PMI. The selection of the optimal method should be based on these factors. The data obtained are critical for planning the preanalytical stage of the study.

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