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Title: Mutational scanning of RB1 gene by multiplex PCR
Authors: Joseph, Biju
Narayana, Komaravelly Siva
Mamatha, Gandra
Raman, Gayathree
Keywords: cost analysis;mPCR;mutations;RB1 gene
Issue Date: Apr-2005
Publisher: CSIR
IPC Code: Int. Cl.7 C12N15/07; 15/10
Abstract:  Multiplex PCR (mPCR) strategy was applied for mutational screening of the exons of RB1 gene in retinoblastoma patients from India. After ethanol purification, the amplified products were directly cycle-sequenced using fluorescent dNTP (deoxy nucleotide triphosphate) ready reaction mix, followed by sequencing in an ABI PRISM 310 automated sequencer. Eighteen of the 27 exons of RB1 gene were amplified in eight multiplex groups with direct savings of 46% of PCR time and 30% of reagents compared to individual amplifications. Thus, the mPCR strategy for sequencing comparatively offered rapid diagnosis and considerable amount of savings in PCR time and reagents’ cost. The mutation results were used for genetic counselling of the patients and their families.
Page(s): 194-200
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.04(2) [April 2005]

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