AGG interruption detection in FMR1 premutation carriers by single-molecule sequencing--run2_repeat
Homo sapiens
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.
ENA first public
2016-11-30
ENA last update
2016-10-13
External Id
SAMEA4378180
INSDC center alias
KU LEUVEN - UNIVERSITY OF LEUVEN
INSDC center name
KU LEUVEN - UNIVERSITY OF LEUVEN
INSDC first public
2016-11-30T17:01:26Z
INSDC last update
2016-10-13T09:54:50Z
INSDC status
public
Submitter Id
AGGdetection_run2.0_repeat
common name
human
sample name
AGGdetection_run2.0_repeat