Comprehensive DNA copy number profile and BAC library construction of an Indian individual |
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| Authors: | Chakrabarty Sanjiban D'Souza Reena R Bellampalli Ravishankara Rotti Harish Saadi Abdul V Gopinath Puthiya M Acharya Raviraja V Govindaraj Periyasamy Thangaraj Kumarasamy Satyamoorthy Kapaettu |
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| Affiliation: | Division of Biotechnology, Manipal Life Sciences Centre, Manipal University, Manipal 576104, Karnataka state, India. sanjiban.c@manipal.edu |
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| Abstract: | Bacterial artificial chromosomes (BACs) are used in genomic variation studies due to their capacity to carry a large insert, their high clonal stability, low rate of chimerism and ease of manipulation. In the present study, an attempt was made to create the first genomic BAC library of an anonymous Indian male (IMBL4) consisting of 100,224 clones covering the human genome more than three times. Restriction mapping of 255 BAC clones by pulse field gel electrophoresis confirmed an average insert size of 120 kb. The library was screened by PCR using SHANK3 (SH3 and multiple ankyrin repeat domains 3) and OLFM3 (olfactomedin 3) specific primers. A selection of clones was analyzed by fluorescent in situ hybridization (FISH) and sequencing. Fine mapping of copy number variable regions by array based comparative genomic hybridization identified 467 CNVRs in the IMBL4 genome. The IMBL4 BAC library represents the first cataloged Indian genome resource for applications in basic and clinical research. |
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| Keywords: | BAC, bacterial artificial chromosome PFGE, pulse field gel electrophoresis CNV, copy number variation arrayCGH, array based comparative genomic hybridization PCR, polymerase chain reaction SHANK3, SH3 and multiple ankyrin repeat domains 3 OLFM3, olfactomedin 3 FISH, fluorescent in situ hybridization |
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