文献类型： 外文期刊
作者： Li, H. 1 ; Li, J. 1 ; Cong, X. H. 2 ; Duan, Y. B. 2 ; Li, L. 2 ; Wei, P. C. 2 ; Lu, X. Z. 2 ; Yang, J. B. 2 ;
作者机构： 1.Chinese Acad Sci, Key Lab Ion Beam Bioengn, Inst Tech Biol & Agr Engn, Hefei, Peoples R China
2.Anhui Acad Agr Sci, Key Lab Rice Genet Breeding Anhui Prov, Rice Res Inst, Hefei, Peoples R China
3.Anhui Agr Univ, Coll Life Sci, Hefei, Peop
关键词： Genomic DNA extraction;Liquid nitrogen-free protocol;High-throughput protocol;Phenol-chloroform-free protocol;High-quality protocol;Real-time PCR
期刊名称：GENETICS AND MOLECULAR RESEARCH （ 影响因子：0.764； 五年影响因子：0.912 ）
ISSN： 1676-5680
年卷期： 2013 年 12 卷 4 期
页码：
收录情况： SCI
摘要： The isolation of high-quality genomic DNA (gDNA) is a crucial technique in plant molecular biology. The quality of gDNA determines the reliability of real-time polymerase chain reaction (PCR) analysis. In this paper, we reported a high-quality gDNA extraction protocol optimized for real-time PCR in a variety of plant species. Performed in a 96-well block, our protocol provides high throughput. Without the need for phenol-chloroform and liquid nitrogen or dry ice, our protocol is safer and more cost-efficient than traditional DNA extraction methods. The method takes 10 mg leaf tissue to yield 5-10 mu g high-quality gDNA. Spectral measurement and electrophoresis were used to demonstrate gDNA purity. The extracted DNA was qualified in a restriction enzyme digestion assay and conventional PCR. The real-time PCR amplification was sufficiently sensitive to detect gDNA at very low concentrations (3 pg/mu L). The standard curve of gDNA dilutions from our phenol-chloroform-free protocol showed better linearity (R-2 = 0.9967) than the phenol-chloroform protocol (R-2 = 0.9876). The results indicate that the gDNA was of high quality and fit for real-time PCR. This safe, high-throughput plant gDNA extraction protocol could be used to isolate high-quality gDNA for real-time PCR and other downstream molecular applications.
- 相关文献
作者其他论文 更多>>
-
Effects of cage stocking density on the production performance, serum biochemistry, immune level, and intestinal morphology of 2 laying hen breeds
作者：Wan, Y.;Guan, H.;Ma, R.;Qi, R.;Li, J.;Liu, W.;Li, Y.;Zhan, K.;Guan, H.;Wang, D.
关键词：chicken; breed; cage stocking density; production performance; intestinal morphology
-
Molecular Cloning and Abiotic Stress Expression Analysis of GTPase Era Gene in Mulberry (Morus alba L.)
作者：Guo, P.;Du, Q. -X.;Zheng, D. -Y.;Dominic, K. -T.;Ackah, M.;Li, Y.;Adolf, A.;Li, L.;Zhao, W. -G.;Hu, F.;Lin, Q.
关键词：Morus alba; GTPase Era; molecular cloning; sequence analysis; expression patterns; abiotic stress
-
Development of SNP-based dCAPS markers for identifying male sterile gene tms5 in two-line hybrid rice
作者：Song, F. S.;Ni, J. L.;Li, L.;Ni, D. H.;Yang, J. B.;Qian, Y. L.
关键词：Marker-assisted selection;tms5 gene;dCAPS;Two-line rice;Purity detection
-
A simplified genomic DNA extraction protocol for pre-germination genotyping in rice
作者：Duan, Y. B.;Zhao, F. L.;Chen, H. D.;Sheng, W.;Teng, J. T.;Zhang, A. M.;Xue, J. P.;Li, H.;Ni, D. H.;Wei, P. C.
关键词：DNA extraction;Half-seed;Polymerase chain reaction;Pre-germination genotyping;Oryza sativa L.
-
High-throughput, low-cost, and event-specific polymerase chain reaction detection of herbicide tolerance in genetically modified soybean A2704-12
作者：Ma, H.;Ma, H.;Li, H.;Li, J.;Wang, X. F.;Wei, P. C.;Li, L.;Yang, J. B.;Li, H.;Wei, P. C.;Li, H.;Li, J.
关键词：Genetically modified soybean A2704-12;Event-specific;Herbicide-tolerance;High-throughput;Qualitative;Quantitative
-
A novel synthetic Cry1Ab gene resists rice insect pests
作者：Song, F. S.;Song, F. S.;Ni, D. H.;Li, H.;Duan, Y. B.;Yang, Y. C.;Ni, J. L.;Lu, X. Z.;Wei, P. C.;Li, L.;Yang, J. B.;Li, H.;Wei, P. C.
关键词：Gene modification;GC content;Synonymous codons;Cry1Ab;Insect resistance
-
Quantitative detection of the rice false smut pathogen Ustilaginoidea virens by real-time PCR
作者：Li, H.;Ni, D. H.;Li, J.;Li, H.;Ni, D. H.;Duan, Y. B.;Li, J.;Song, F. S.;Li, L.;Wei, P. C.;Yang, J. B.;Li, H.;Wei, P. C.;Duan, Y. B.;Song, F. S.;Chen, Y.
关键词：Rice false smut;Real-time PCR;Detection;Quantification;Ustilaginoidea virens
