Journal: Biology Methods & Protocols
Article Title: PrimerPooler: automated primer pooling to prepare library for targeted sequencing
doi: 10.1093/biomethods/bpx006
Figure Lengend Snippet: Coverage of targeted sequence regions by Fluidigm Access Array multiplex PCR and Illumina MiSeQ sequencing. Two FFPE DNA samples were tested under various experimental conditions including 50- and 75-ng DNA for pre-amplication, and 1l of 2.5- and 5.0-fold diluted pre-amplified products for Fluidigm Access Array multiplex PCR. An average of 3269 reads was achieved for the targeted regions, with 95% of targets covered by at least 50 reads, the minimal depth of reads for confident variant calling.
Article Snippet: Based on the two DNA samples tested, both 50ng and 75ng starting amounts of DNA yielded very similar results, and similarly there was no obvious benefit when less diluted pre-amplified product (2.5 versus 5-fold dilution) was used for Fluidigm Access Array multiplex PCR ( ).
Techniques: Sequencing, Multiplex Assay, Amplification, Variant Assay