Comparison of three genomic DNA extraction methods to obtain high DNA quality from maize

被引:135
作者
Abdel-Latif, Amani [1 ]
Osman, Gamal [2 ,3 ]
机构
[1] Univ Alexandria, Dept Bot & Microbiol, Fac Sci, Alexandria, Egypt
[2] Umm Al Qura Univ, Dept Biol, Fac Sci Appl, POB 715, Mecca 21955, Saudi Arabia
[3] Agr Genet Engn Res Inst, Giza 12619, Egypt
来源
PLANT METHODS | 2017年 / 13卷
关键词
Internal transcribed spacer (ITS); Internal transcribed spacer 2 (ITS2); DNA extraction; PROTOCOL; PLANTS; FRESH; RAPD;
D O I
10.1186/s13007-016-0152-4
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Background: The world's top three cereals, based on their monetary value, are rice, wheat, and corn. In cereal crops, DNA extraction is difficult owing to rigid non-cellulose components in the cell wall of leaves and high starch and protein content in grains. The advanced techniques in molecular biology require pure and quick extraction of DNA. The majority of existing DNA extraction methods rely on long incubation and multiple precipitations or commercially available kits to produce contaminant-free high molecular weight DNA. Results: In this study, we compared three different methods used for the isolation of high-quality genomic DNA from the grains of cereal crop, Zea mays, with minor modifications. The DNA from the grains of two maize hybrids, M10 and M321, was extracted using extraction methods DNeasy Qiagen Plant Mini Kit, CTAB-method (with/ without 1% PVP) and modified Mericon extraction. Genes coding for 45S ribosomal RNA are organized in tandem arrays of up to several thousand copies and contain codes for 18S, 5.8S and 26S rRNA units separated by internal transcribed spacers ITS1 and ITS2. While the rRNA units are evolutionary conserved, ITS regions show high level of interspecific divergence and have been used frequently in genetic diversity and phylogenetic studies. In this study, the genomic DNA was then amplified with PCR using primers specific for ITS gene. PCR products were then visualized on agarose gel. Conclusion: The modified Mericon extraction method was found to be the most efficient DNA extraction method, capable to provide high DNA yields with better quality, affordable cost and less time.
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页数:9
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共 28 条
[1]  
Abdulla M, 2010, AUST J CROP SCI, V4, P23
[2]   Development of transgenic wheat (Triticum aestivum L.) expressing avidin gene conferring resistance to stored product insects [J].
Abouseadaa, Heba H. ;
Osman, Gamal H. ;
Ramadan, Ahmed M. ;
Hassanein, Sameh E. ;
Abdelsattar, Mohamed T. ;
Morsy, Yasser B. ;
Alameldin, Hussien F. ;
El-Ghareeb, Doaa K. ;
Nour-Eldin, Hanan A. ;
Salem, Reda ;
Gad, Adel A. ;
Elkhodary, Soheir E. ;
Shehata, Maher M. ;
Mahfouz, Hala M. ;
Eissa, Hala F. ;
Bahieldin, Ahmed .
BMC PLANT BIOLOGY, 2015, 15
[3]   Removal of PCR inhibitors from soil DNA by chemical flocculation [J].
Braid, MD ;
Daniels, LM ;
Kitts, CL .
JOURNAL OF MICROBIOLOGICAL METHODS, 2003, 52 (03) :389-393
[4]   Barcoding the kingdom Plantae: new PCR primers for ITS regions of plants with improved universality and specificity [J].
Cheng, Tao ;
Xu, Chao ;
Lei, Li ;
Li, Changhao ;
Zhang, Yu ;
Zhou, Shiliang .
MOLECULAR ECOLOGY RESOURCES, 2016, 16 (01) :138-149
[5]   PRE-GERMINATION GENOTYPIC SCREENING USING PCR AMPLIFICATION OF HALF-SEEDS [J].
CHUNWONGSE, J ;
MARTIN, GB ;
TANKSLEY, SD .
THEORETICAL AND APPLIED GENETICS, 1993, 86 (06) :694-698
[6]   New multiplex PCR methods for rapid screening of genetically modified organisms in foods [J].
Datukishvili, Nelly ;
Kutateladze, Tamara ;
Gabriadze, Inga ;
Bitskinashvili, Kakha ;
Vishnepolsky, Boris .
FRONTIERS IN MICROBIOLOGY, 2015, 6
[7]   Comparative evaluation of DNA extraction methods for food crops [J].
Datukishvili, Nelly ;
Gabriadze, Inga ;
Kutateladze, Tamara ;
Karseladze, Marina ;
Vishnepolsky, Boris .
INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY, 2010, 45 (06) :1316-1320
[8]   Isolation, cloning, and overexpression of vip3Aa gene isolated from a local Bacillus thuringiensis [J].
El-Ghareeb, D. K. ;
Osman, G. H. ;
El Baz, A. F. .
BIOCONTROL SCIENCE AND TECHNOLOGY, 2012, 22 (01) :11-21
[9]  
Gabriadze I., 2014, Int. J. Food Sci. Nutr, V3, P199, DOI [10.11648/j.ijnfs.20140303.21, DOI 10.11648/J.IJNFS.20140303.21]
[10]   Evaluation of extraction methodologies for corn kernel (Zea mays) DNA for detection of trace amounts of biotechnology-derived DNA [J].
Holden, MJ ;
Blasic, JR ;
Bussjaeger, L ;
Kao, C ;
Shokere, LA ;
Kendall, DC ;
Freese, L ;
Jenkins, GR .
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 2003, 51 (09) :2468-2474