An improved method for genomic DNA extraction from strawberry leaves

被引：33

作者：

Nunes, Claudineia Ferreira ^{[1
]}

Ferreira, Juliano Lino ^{[1
]}

Nunes Fernandes, Monique Carolina ^{[1
]}

Breves, Samera de Souza ^{[1
]}

Generoso, Andressa Leal ^{[1
]}

Fontes Soares, Barbara Dantas ^{[1
]}

Carvalho Dias, Mario Sergio ^{[1
]}

Pasqual, Moacir ^{[2
]}

Borem, Aluizio ^{[3
]}

de Almeida Cancado, Geraldo Magela ^{[1
]}

机构：

[1] Empresa Pesquisa Agr Minas Gerais EPAMIG, Plant Biotechnol Lab, BR-37780000 Caldas, MG, Brazil

[2] Univ Fed Lavras UFLA, Dept Agr, Lavras, MG, Brazil

[3] Univ Fed Vicosa, Dept Agr, Vicosa, MG, Brazil

来源：

CIENCIA RURAL | 2011年 / 41卷 / 08期

关键词：

Fragaria ananassa; genomic DNA; polivinilpirrolidone; lyophilization; PCR;

D O I：

10.1590/S0103-84782011000800014

中图分类号：

S3 [农学（农艺学）];

学科分类号：

0901 ;

摘要：

Several extraction methods of genomic DNA for identification and characterization of genetic diversity in different plant species are routinely applied during molecular analysis. However, the presence of undesirable compounds such as polyphenols and polysaccharides is one of the biggest problems faced during the isolation and purification of high quality DNA in plants. Therefore, achievement of fast and accurate methods for DNA extraction is crucial in order to produce pure samples. Leaves of strawberry genotypes (Fragaria ananassa) have high contents of polysaccharides and polyphenols which increase the sample viscosity and decrease the DNA quality, interfering with the PCR performance. Thereby, in this study we evaluated the quality and amount of genomic DNA extracted from young leaves of strawberry after tissue lyophilization and maceration in presence of polivinilpirrolidone (PVP). The CTAB method was used as reference procedure and it was modified to improve the DNA extraction. The modifications consisted of tissue lyophilization overnight until it was completely freeze-dried and addition of PVP during the tissue maceration in liquid nitrogen. The results showed the efficiency and reliability of the modified method compared to the unmodified method, indicating that combination of lyophilization and PVP improve the quality and amount of the DNA extracted from strawberry leaves.

引用

页码：1383 / 1389

页数：7

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