Modified salting-out method: High-yield, high-quality genomic DNA extraction from whole blood using laundry detergent

被引:115
|
作者
Nasiri, H [1 ]
Forouzandeh, M [1 ]
Rasaee, MJ [1 ]
Rahbarizadeh, F [1 ]
机构
[1] Tarbiat Modarres Univ, Dept Med Biotechnol, Sch Med Sci, Tehran, Iran
关键词
DNA extraction; salting-out; polymerase chain reaction; genomic DNA; laundry detergent;
D O I
10.1002/jcla.20083
中图分类号
R446 [实验室诊断]; R-33 [实验医学、医学实验];
学科分类号
1001 ;
摘要
Different approaches have been used to extract DNA from whole blood. In most of these methods enzymes (such as proteinase K and RNAse A) or toxic organic solvents (such as phenol or guanidine isothiocyanate) are used. Since these enzymes are expensive, and most of the materials that are used routinely are toxic, it is desirable to apply an efficient DNA extraction procedure that does not require the use of such materials. In this study, genomic DNA was extracted by the salting-out method, but instead of using an analytical-grade enzyme and chemical detergents, as normally used for DNA isolation, a common laundry powder was used. Different concentrations of the powder were tested, and proteins were precipitated by NaCl-saturated distilled water. Finally, DNA precipitation was performed with the use of 96% ethanol. From the results, we conclude that the optimum concentration of laundry powder for the highest yield and purity isolated DNA is 30 mg/mL. The was optimized, and a final protocol suggested. Following the same DNA was extracted from 100 blood samples, and their amounts were found to >50 mu g/mL of whole blood. The integrity the DNA fragments was confirmed agarose gel electrophoresis. the extracted DNA was used as a for PCR reaction. The results obtained PCR showed that the final solutions extracted DNA did not contain any material for the enzyme used in the reaction, and indicated that the DNA was of good quality. These show that this method is simple, fast, and cost-effective, and can be used medical la- boratories and research centres.
引用
收藏
页码:229 / 232
页数:4
相关论文
共 50 条
  • [41] HIGH-THROUGHPUT GENOMIC DNA EXTRACTION FROM BLOOD SAMPLE USING MAGSTATION SYSTEM
    Liu Hongna
    Li Song
    Tian Lan
    Li Zhiyang
    Ma Chao
    He Nongyue
    IFPT'6: PROGRESS ON POST-GENOME TECHNOLOGIES, PROCEEDINGS, 2009, : 84 - 86
  • [42] Development of a High-Quality/Yield Long-Read Sequencing-Adaptable DNA Extraction Method for Crop Seeds
    Shioya, Naohiro
    Ogiso-Tanaka, Eri
    Watanabe, Masanori
    Anai, Toyoaki
    Hoshino, Tomoki
    PLANTS-BASEL, 2023, 12 (16):
  • [43] Modified low-salt CTAB extraction of high-quality DNA from contaminant-rich tissues
    Arseneau, Jean-Rene
    Steeves, Royce
    Laflamme, Mark
    MOLECULAR ECOLOGY RESOURCES, 2017, 17 (04) : 686 - 693
  • [44] Improved extraction and recrystallization of high-quality aleuritic acid from natural resins using a modified technique
    Ali, Mohd
    Hazra, Dipak Kumar
    Kumar, Yengkhom Bijen
    Karmakar, Rajab
    SEPARATION SCIENCE AND TECHNOLOGY, 2022, 57 (18) : 2916 - 2922
  • [45] An optimized mini-preparation method to obtain high-quality genomic DNA from mature leaves of sunflower
    Li, J. T.
    Yang, J.
    Chen, D. C.
    Zhang, X. L.
    Tang, Z. S.
    GENETICS AND MOLECULAR RESEARCH, 2007, 6 (04) : 1064 - 1071
  • [46] A Modified SDS-Based DNA Extraction Method for High Quality Environmental DNA from Seafloor Environments
    Natarajan, Vengadesh Perumal
    Zhang, Xinxu
    Morono, Yuki
    Inagaki, Fumio
    Wang, Fengping
    FRONTIERS IN MICROBIOLOGY, 2016, 7
  • [47] Research note: methodology for high-quality RNA extraction from poultry whole blood for further gene expression analysis
    Mewis, J. L.
    Sun, X.
    Zuidhof, M. J.
    Guan, L. L.
    BRITISH POULTRY SCIENCE, 2014, 55 (02) : 194 - 196
  • [48] Improved protocol for extracting genomic DNA from frozen formalin-fixed tissue resulting in high-quality whole mtDNA
    Wang, H. W.
    Xu, Y.
    Zhang, H. F.
    Zeng, Y. J.
    Ren, L.
    Miao, Y. L.
    Luo, H. Y.
    Wang, K. H.
    GENETICS AND MOLECULAR RESEARCH, 2016, 15 (03)
  • [49] Efficient isolation method for high-quality genomic DNA from cicada exuviae (vol 7, pg 8161, 2017)
    Hoa Quynh Nguyen
    Kim, Ye Inn
    Borzee, Amael
    Jang, Yikweon
    ECOLOGY AND EVOLUTION, 2018, 8 (06): : 3570 - 3570
  • [50] High-Quality and -Quantity DNA Extraction from Frozen Archival Blood Clots for Genotyping of Single-Nucleotide Polymorphisms
    Bank, Steffen
    Nexo, Bjorn Andersen
    Andersen, Vibeke
    Vogel, Ulla
    Andersen, Paal Skytt
    GENETIC TESTING AND MOLECULAR BIOMARKERS, 2013, 17 (06) : 501 - 503