An improved method for efficient recovery of high quality DNA from date palm (Phoenix dactylifera L; Arecaceae)

Abstract

Date palm ( Phoenix dactylifera L; Arecaceae ) is one of a few fruit trees that can remarkably grow in dessert agroecosystems that are characterized by extreme temperature fluctuations. Due to increasing demands for dates in the global market and commercial cultivation in many countries, the tree is currently under extensive research in many countries, particularly to improve the germplasm using different molecular tools. Most molecular techniques largely depend on good quality DNA in significant quantities, which are highly compromised by the presence of various contaminants in DNA. The traditional cetyltrimethylammoniumbromide (CTAB) based method has failed to produce good quality DNA from date palm due to hard fibrous nature of tissue. On the basis of previous studies, commercial DNA extraction kits are not economical although they are very effective. Therefore, we have developed an improved DNA extraction protocol by modifying the original CTAB method to produce extra pure DNA in large quantities. The novel method has been validated using different quality testing approaches. This cost-effective method can be used successfully for DNA extraction from date palm. Moreover, this improved method may have potential for DNA extraction from other palms that have similar leaf texture to date palm leaves, but this method needs to be tested for other palms before being used. The improved method has following key modifications: Grinding of plant tissue in liquid nitrogen and subsequent lysis of cells in CTAB buffer that has increased concentration of ß-mercaptoethanol Repeated steps of chloroform: IAA extraction and ethanol washing Addition of RNase A before the DNA precipitation step

The sweet potato B-box transcription factor gene IbBBX28 negatively regulates drought tolerance in transgenic Arabidopsis

“…The Open Reading Frameb (ORF) sequence of IbBBX28 (GenBank accession number OP047916) was previously cloned in our lab, and the gene structure of the genomic DNA sequence of IbBBX28 was predicted from the database ( https://121.36.193.159/blast.html ). The genomic DNA was extracted from XZ-3 root sample using the CTAB method ( Safeena et al, 2021 ). Using the genomic DNA as template, the genomic DNA sequence and promoter sequence of IbBBX28 were cloned by PCR.…”

“…36.193.159/blast.html). The genomic DNA was extracted from XZ-3 root sample using the CTAB method (Safeena et al, 2021). Using the genomic DNA as template, the genomic DNA sequence and promoter sequence of IbBBX28 were cloned by PCR.…”

Evaluating the genetic diversity of Erythropalum scandens based on using inter-simple sequence repeat markers

“…Genomic DNA was extracted using a modi ed CTAB method (Safeena et al 2021), and its quality was examined using 0.8% agarose gel electrophoresis. The concentration and purity of DNA were determined using an ultra-micro UV-Vis spectrophotometer (Thermo NanoDrop One, America).…”

Optimización de extracción de ADN de polen apícola para caracterización molecular

“…Un punto clave para obtener exitosamente códigos de barras de ADN, es la extracción del material genético, donde se requiere un método rápido, fácil y económico, del cual resulte un alto rendimiento y calidad de ADN. Para garantizar la reproducibilidad de los resultados es necesario descartar la presencia de inhibidores de la PCR (Safeena et al 2021). Un ADN libre de inhibidores permite realizar no solo la amplificación de los códigos de barras, si no también hace posible la detección de polen de plantas transgénicas, alergénicas o bien de plantas invasoras (Guertler et al 2014).…”