Chinese Journal of Nephrology, Dialysis & Transplantation ›› 2018, Vol. 27 ›› Issue (3): 245-249.DOI: 10.3969/j.issn.1006-298X.2018.03.009
• Article • Previous Articles Next Articles
Online:
Published:
Abstract:
Objective:To investigate the quality and yield of DNA extracted from blood cell after longterm(more than 10 years) storage at -80℃. Methodology:The 150 blood cell samples enrolled in the study were selected from the Renal Biobank of National Clinical Research Center of Kidney Diseases,which were isolated from 3 ml anticoagulant blood having been preserved for more than 10 years.Extraction of DNA was performed on automatic nucleic acid extraction instrument by magnetic beads method.Assay of the purity and yield of DNA by Nanodrop and the integrity by agarose gel electrophoresis. Results:Of the DNA extracted from 150 blood cell sampels,the mean value of A260/A280 is 184,A260/A230 is 227 and yield is 18757 μgAgarose gel electrophoresis showed that all the DNA bands were relatively concentrated without dispersion distribution,and DNA molecular weight larger than 20 kb.In the 150 DNA samples,there are 134 DNA 17≤A260/A280≤19,accounting for 893%,120 DNA A260/A230≥20,accounting for 80%,115 DNA 17≤A260/A280≤19 and A260/A230≥20,accounting for 767%.The yield of 143 DNA ≥10 μg,accounting for 953%.Of which,there are 132 DNA yield≥10 μg and 17≤A260/A280≤19,accounting for 88%.Further,114 DNA yield≥10 μg,17≤A260/A280≤19,and A260/A230≥20,accounting for 76%. Conclusion: DNA extracted from blood cell samples by magnetic beads mostly remains high quality and quantity after longterm cryopreservation in our center,which can meet the demand of downstream experiments.This research offers a reference for selecting samples for scientific research that had been preserved for more than 10 years in Biobank.
Key words: blood cell samples, long-termcryopreservation, DNA
DOU Weijuan,LU Yinghui,TANG Rong,et al. Quality and yield of DNA extracted from blood cell after longterm cryopreservation[J]. Chinese Journal of Nephrology, Dialysis & Transplantation, 2018, 27(3): 245-249.
0 / / Recommend
Add to citation manager EndNote|Ris|BibTeX
URL: http://www.njcndt.com/EN/10.3969/j.issn.1006-298X.2018.03.009
http://www.njcndt.com/EN/Y2018/V27/I3/245