Hi all,
I'm new to SEQanswers and a PhD student at Ohio State. A faculty member recently purchased a PCR amplicon sequencing kit from Illumina and is allowing me to piggyback on his upcoming sequencing run. While pursuing my Master's, I used ISSR primers to look at clonality in a species of sea anemone, and this approach would generate anywhere from 5-15 bands/primer. I was thinking that I might be able to take PCR product from these ISSR primers, ligate the Illumina primers/barcodes on the PCR amplicons, and sequence them using this new kit. Has anyone done this before?
It was suggested I post this question here since he hadn't heard of anyone doing this before. He also suggested that the repeats might cause secondary structure during sequencing and may mess up the run... Any thoughts?
I appreciate the insight!
Ben
I'm new to SEQanswers and a PhD student at Ohio State. A faculty member recently purchased a PCR amplicon sequencing kit from Illumina and is allowing me to piggyback on his upcoming sequencing run. While pursuing my Master's, I used ISSR primers to look at clonality in a species of sea anemone, and this approach would generate anywhere from 5-15 bands/primer. I was thinking that I might be able to take PCR product from these ISSR primers, ligate the Illumina primers/barcodes on the PCR amplicons, and sequence them using this new kit. Has anyone done this before?
It was suggested I post this question here since he hadn't heard of anyone doing this before. He also suggested that the repeats might cause secondary structure during sequencing and may mess up the run... Any thoughts?
I appreciate the insight!
Ben