A simple method for DNA extraction from mature date palm leaves: impact of sand grinding and composition of lysis buffer.
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.
Molecular Fingerprinting and Biodiversity Unit, Prince Sultan Research Chair for Environment and Wildlife, College of Sciences, King Saud University, Riyadh, Saudi Arabia; E-Mails: iaarif@hotmail.com ; (I.A.A.); mabakir@yahoo.com (M.A.B.); nanisahamed@rediffmail.com (A.A.); alfarhan@ksu.edu.sa (A.H.A.F.); homaidan@ksu.edu.sa (A.A.A.H.); msadoon@ksu.edu.sa (M.A.S.); abahkali@ksu.edu.sa (A.H.B.); mshobrak@gmail.com (M.S.).
Ibrahim A
IA
Arif
Mohammad A
MA
Bakir
Haseeb A
HA
Khan
Anis
A
Ahamed
Ahmad H
AH
Al Farhan
Ali A
AA
Al Homaidan
Mohammad
M
Al Sadoon
Ali H
AH
Bahkali
Mohammad
M
Shobrak
D028023:Arecaceae / Q000737:chemistry*; D002458:Cell Fractionation / Q000379:methods; D005591:Chemical Fractionation / Q000379:methods; D018744:DNA, Plant / Q000737:chemistry / Q000302:isolation & purification*; D006868:Hydrolysis; D018515:Plant Leaves / Q000737:chemistry*
D018744:DNA, Plant