Figure 1
From: Targeted next-generation sequencing for routine clinical screening of mutations
Outline of the tSMS gene capture approach. Tumor material can be collected and (a) snap frozen to preserve intact DNA and RNA or (b) fixed in formalin then embedded in paraffin for section and histopathological review. Following (c) DNA extraction, (d) the DNA is broken by sonication to approximately 200 bp fragments. Green, sequences from the BRCA1 region; black, other regions. Formalin fixed material provides fragmented DNA, removing the need for a sonication step. (e) DNA fragments are hybridized to the flow cell, which is covered with oligonucleotides with sequence complementary to the region corresponding to the BRCA1 gene. BRCA1 sequences are therefore enriched on the flow cell. (f) Each individual molecule of DNA is then sequenced simultaneously by sequential addition of fluorescently labeled nucleotides (purple).