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Original Articles Open Access
Phosphoinositide-3- kinases (PI3Ks) are widely involved in cellular processes and can phosphorylate the 3-hydroxyl position of the inositol ring of phosphoinositides. Enhancement of P13Ks functions in cellular processes cannot be overlooked in protein interactions and signal trafficking. The effect of magnesium ions on phosphoinositide-3- kinase C2β C2 domain was aimed to be investigated. The amplification of a DNA fragment of PI3K C2β C2 domain using polymerase chain reaction (PCR) technique was employed. The oligonucleotide primers (anti sense, sense) were mixed with buffer, dNTPs, vent DNA polymerase and variable concentration (500, 1000 and 1500μM) of magnesium sulphate. The PCR products were analysed using agarose gel electrophoresis and purified with using QIAquick PCR purification kit to determine the effect of Magnesium (Mg2+) concentration on the amplified gene product for gene manipulation and purification of the amplified gene product. The successful qualitative PCR analysis of human PI3K C2β C2 domain in the absence of an additional magnesium ions yielded the specific amplicon of the desired product size below 4000bp and PCR products were successfully purified to remove debris from the products. The purified PCR products can be used for cloning.
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Author(s): Banigo Alma Tamunonengiofori and Azeez Taofik Oladimeji
Magnesium ions, Phosphoinositide-3-Kinase (PI3K), PI3K C2β C2 domain, PCR technique, Magnesium ions