Abstract
Extraction of DNA in many plants is difficult because of the presence of metabolites that interfere with DNA isolation procedures and downstream applications such as DNA restriction, replications, amplification, as well as cloning. Modified procedure based on the hexadecyltrimethyl ammoniumbromide (CTAB) method is used to isolate DNA from tissues containing high levels of polysaccharides. The procedure is applicable to both ripped and unripe fruits of Annona senegalensis . This modified CTAB (2%) protocol include the use of 1.4 M NaCl, 1% polyvinylpyrrolidine (PVP), 1% mercaptoethanol and 100% absolute ethanol in the extraction as well as reducing the centrifugation times during the separation and precipitation of the DNA. This method solved the problems of DNA contamination, degradation and low yield due to binding or co-precipitation with starches. The isolated DNA proved amenable to polymerase chain reaction (PCR) amplification and restriction digestion. This technique is fast, reproducible, and can be applied for simple sequence repeats (SSR)-PCR markers identification. Key words : Annona senegalensis , genomic DNA, fruits, modified, markers.
Highlights
Annona senegalensis Pers. is a member of the Annonaceae family and it is a species of seed vegetable which grow both on dry and raining seasons
To help in minimizing time spent doing this step, 1 ml of 2% CTAB was measured in a 2 ml micro centrifuge tube to which 100 μl of mercaptoethanol (1%, v/v) was added and the tube placed in a 60°C water bath until ready for use
Addition of the pre-warmed, pre-measured CTAB buffer to the frozen leaf tissue contained in the prechilled conical tube saves precious time in bringing the tissue from -80 to 60°C as rapidly as possible resulting in 1% mercaptoethanol produced nucleic acid pellets that were not nearly brown
Summary
Annona senegalensis Pers. is a member of the Annonaceae family and it is a species of seed vegetable which grow both on dry and raining seasons. Is a member of the Annonaceae family and it is a species of seed vegetable which grow both on dry and raining seasons. It is a savannah plant which is widely spread from Senegal to Nigeria, in Central African Republic (Abdullahi et al, 2012). It produces seeds which are ovate in shape, very small in size and open by mechanical explosion. A. senegalensis is common in Southern part and in Niger State of Nigeria; where they use the seeds and fruits in making soup (soup harder). The neglect of some local vegetables coupled with the growing reduction in their consumption prompted this research
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