A Family Based Whole Exome Sequence Study to Indentify Modifier Genes for Phenotype Heterogeneity Between Severe and Non-Severe Thalassemia Patients

* Corresponding author: Prof. Anupam Basu, Department of Zoology, The University of Burdwan, PurboBarddhaman, West Bengal- 713104, India, Email- abasu@zoo.buruniv.ac.ina & b are equal contribution Whole Expanded Exome Sequencing Study of two families father, mother and index cases (trio) was undertaken for two E/ beta thalassemia subjects with same HBB genotype. Approximately 200ng of DNA was taken from each individual and shared into 300-400bp fragment. Then the shared fragments are end repair. Klenow exonuclease was used to add an adapter. After adapter ligation 10 cycle PCR amplification was done for each sample. The targeted Exome was captured by the Agilent Sure select XT Human all Exome V6+ UTR kit as per the manufacturer’s protocol. Captured library was then amplified 10 cycles with 8 bp index sequence for each sample. Then the indexed capture library was pooled together. Pair end sequencing of the pooled library was performed in Illumina HiSeq2500 using Illumina HiSeq SBS kit. Finally, both the genes, inherited and denovo, from both the subjects were separately functionally annotated by DAVID online tools 6.8. Functionally annotation result shows that in case of subject-1, 6 KEGG pathway were involved. These are Adherent junction, Protein digestion and absorption, Inflamatory Bowel Disease, Amoebiasis, PPAR signaling pathway and glycolysis or gluconeogenesis. Interestingly in case of subject-2, only 2 KEGG pathway were found, Thyroid hormone synthesis and carbon metabolism.