A simple method for DNA extraction from mature date palm leaves: impact of sand grinding and composition of lysis buffer.
A simple method for DNA extraction from mature date palm leaves: impact of sand grinding and composition of lysis buffer.
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DOI：
10.3390/ijms11093149
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发表时间：
2010-09-08
影响因子：
5.6
通讯作者：
Shobrak M
中科院分区：
文献类型：
--
作者：
Arif IA;Bakir MA;Khan HA;Ahamed A;Al Farhan AH;Al Homaidan AA;Al Sadoon M;Bahkali AH;Shobrak M
分子标记技术已被广泛应用于枣（Phoenix dactylifera L.）槟榔科）与生物多样性保护。高纯度DNA的分离是PCR扩增和随后使用的先决条件，例如最近开发用于条形码的基因的DNA指纹图谱和测序。为了避免与液氮的保存和使用有关的问题，我们研究了用于研磨枣椰叶的无菌砂。在这项研究中，氯化钠（NaCl），聚乙烯吡咯烷酮（PVP）和氯化锂（LiCl）与十六烷基三甲基溴化铵（CTAB）的DNA产量足够的纯度和PCR扩增的方法的单独和组合的影响进行了评价。与加入NaCl相比，LiCl和PVP单独或一起存在于裂解缓冲液中并没有显著提高DNA产率和纯度。我们的研究表明，用无菌沙研磨椰枣叶并在裂解缓冲液中加入NaCl（1.4 M），而不使用昂贵的液氮、PVP和LiCl，提供了足够纯度的DNA产量，适用于PCR扩增。
Molecular marker techniques have been widely used for cultivar identification of inbred date palms (Phoenix dactylifera L.; Arecaceae) and biodiversity conservation. Isolation of highly pure DNA is the prerequisite for PCR amplification and subsequent use such as DNA fingerprinting and sequencing of genes that have recently been developed for barcoding. To avoid problems related to the preservation and use of liquid nitrogen, we examined sterile sand for grinding the date palm leaves. Individual and combined effects of sodium chloride (NaCl), polyvinylpyrrolidone (PVP) and lithium chloride (LiCl) with the cetyltrimethylammonium bromide (CTAB) method for a DNA yield of sufficient purity and PCR amplification were evaluated in this study. Presence of LiCl and PVP alone or together in the lysis buffer did not significantly improve the DNA yield and purity compared with the addition of NaCl. Our study suggested that grinding of date palm leaf with sterile sand and inclusion of NaCl (1.4 M) in the lysis buffer without the costly use of liquid nitrogen, PVP and LiCl, provides a DNA yield of sufficient purity, suitable for PCR amplification.
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