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|Title: ||Mutational scanning of RB1 gene by multiplex PCR|
|Authors: ||Joseph, Biju|
Narayana, Komaravelly Siva
|Keywords: ||cost analysis;mPCR;mutations;RB1 gene|
|Issue Date: ||Apr-2005|
|IPC Code: ||Int. Cl.7 C12N15/07; 15/10|
(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.
|ISSN: ||0975-0967 (Online); 0972-5849 (Print)|
|Appears in Collections:||IJBT Vol.04(2) [April 2005]|
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