Patent ID: 6063567
Filing Date: 2000-05-16
Classification: C12Q

Abstract:
A method for identifying mutations in a sample retinoblastoma gene comprising the steps of:(a) quantitatively coamplifying a plurality of exons of the sample retinoblastoma gene using primers complementary to intron regions immediately flanking each of the plurality of exons;(b) determining the lengths of the amplification products for each amplified sample exon and comparing that length to the length of amplification products obtained when a wild-type retinoblastoma gene is amplified using the same primers, whereby differences in length between an amplified sample exon and the corresponding amplified wild-type exon reflect the occurrence on an insertion or deletion mutation in the sample retinoblastoma gene; and(c) determining the nucleic acid sequence of each exon identified in step (b) to contain an insertion or deletion mutation, or in the event no insertion or deletion mutations are identified, determining the nucleic acid sequence of at least one exon of the retinoblastoma gene, sufficient exons being sequenced to identify the presence of a mutation if one exists in the sample retinoblastoma gene, wherein in step (a) exons 4, 5, 7, 8, 10, 11, 19 and 25 are coamplified in a single reaction.