Effects of Centrifugation at Different Levels of Freeze-Thawed Blood on DNA Isolation

Authors : Mevlut Arslan

Pages : 154-163

Doi:10.53433/yyufbed.1130525

View : 13 | Download : 7

Publication Date : 2023-04-30

Article Type : Research Article

Abstract :DNA isolation from blood is a commonly used application to obtain nDNA and mtDNA. It was previously shown that DNA isolation could be performed from the pellet obtained after centrifugation of freeze-thawed blood (FTB), and this pretreatment had constructive results on DNA isolation. However, which centrifugation levels can be used for this pretreatment, and their effects are unknown. The aim of the study was to determine appropriate centrifugation levels for this pretreatment and show their effects on isolated DNA. For this purpose, DNA isolations were carried out from pellet and supernatant obtained by centrifugation at different levels of FTB. Then, spectrophotometric, gel electrophoresis, and real-time PCR analyses were performed in the isolated DNA samples. As a result, centrifugation of FTB at 5,000×g for 2 min or over let genetic material to pellet completely. This also caused to obtain high amount of DNA. mtDNA/nDNA ratios did not change in the isolated DNA samples from pellets obtained by defined centrifugation levels, but the DNA integrity decreased. To conclude, centrifugation of FTB at 5,000×g for 2 min or over can be used to harvest and wash genetic material found in FTB before DNA isolations.
Keywords : DNA izolasyonu, Donmuş-çözünmüş kan, Kan, Santrifüj