Rehan et al. Pakhtunkhwa J. Life Sci. Volume 03, Issue 01-02, 2015, P 11-21 Available online at: www.awkum.edu.pk/PJLS 11 Epigenetic Analysis of OCT4 Gene in Blood Cells of Normal and Leukemic Patients Touseef Rehan 1* , Riffat Tahira 2 , Tabassum Rehan 3 , Ayesha Bibi 4 , Amir Ali Khan 1 1 Department of Biochemistry and Molecular biology, Quaid-i-Azam University, Islamabad, Pakistan 2 Plants Genetic Resources Program, National Agriculture Research Center Islamabad, Pakistan 3 Department of Biotechnolgy, Quaid-i-Azam University, Islamabad, Pakistan 4 Department of Genetics, Hazara University, Mansehra, Pakistan *Corresponding Author: touseefbiochem@gmail.com ABSTRACT Epigenetic changes are one of the many causes of cancer. Our research interest lie in the alteration of methylation in the CG in the promotor of OCT4 in the DNA of ALL(acute lyphoblastic Leukaemia). We examined the alteration of methylation of CG at three CCGG loci in the promotor of Oct4 by restriction digestion of CCGG using MSPI and HPaII followed by PCR to amplify the region flanking the CCGG. This simple yet effective methology was used to assess the alteration of methylation in the CG of specific CCGG in the 60 control and 50 ALL patients. CCGG sequence at 2 loci was selected in the 2.7 Kb upstream regulatory region of Oct4 gene. The DNA from the ALL patients were digested with MSPI and HpaII followed by PCR using the primers that flank the CCGG. The PCR bands were observed in the control while they were absent in the DNA of ALL patients indicating the alteration of methylation at these CGs. Our results demonstrate that the specific CG in the CCGG inside the promoter of OCT4 undergoes demthylation in the ALL patients compared to normal. However, research is needed to confirm our preliminary findings. Key words: Alteration of methylation, Leukaemia, digestion of CCGG, PCR