title: AGG interruption detection in FMR1 premutation carriers by single-molecule sequencing--run2_repeat
organism: Homo sapiens
description: The FMR1 gene contains an unstable CGG repeat in its 5' untranslated region. A repeat with 55-200 CGG units is called a premutation allele and often expands to a full mutation during female germline transmission giving rise to the fragile X syndrome. The risk for a premutation to expand depends on both size and AGG interruptions in the CGG repeat. Unfortunately, the detection of AGG interruptions is hampered by technical difficulties. In order to circumvent the various limitations of TP-PCR, we explored single-molecule sequencing to determine the number and location of AGG interruptions in both males and females carrying FMR premutation alleles. Therefore we first pooled different FMR1 amplicons by using barcodes from PacBio's 96 Barcode design (details can be found in the manuscript) followed by single-molecule sequencing and selection of reads-of-inserts with a minimum of 10 full passes. We demonstrated that single-molecule sequencing enables the determination of not only the repeat size, but also the complete repeat structure including AGG interruptions in male and female alleles with repeats ranging from 45 to 100 CGG units.
attributes.tag: ENA first public
attributes.value: 2016-11-30
attributes.tag: ENA last update
attributes.value: 2016-10-13
attributes.tag: External Id
attributes.value: SAMEA4378180
attributes.tag: INSDC center alias
attributes.value: KU LEUVEN - UNIVERSITY OF LEUVEN
attributes.tag: INSDC center name
attributes.value: KU LEUVEN - UNIVERSITY OF LEUVEN
attributes.tag: INSDC first public
attributes.value: 2016-11-30T17:01:26Z
attributes.tag: INSDC last update
attributes.value: 2016-10-13T09:54:50Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: AGGdetection_run2.0_repeat
attributes.tag: common name
attributes.value: human
attributes.tag: sample name
attributes.value: AGGdetection_run2.0_repeat