Hi,
As shown in Lennon et al (2010, Genome Biol.) and Rodrigue et al (2010, Plos One), the Broad and many other folks seem to routinely employ a double AMPure bead approach to obviate the need for gels and to make size selection more scalable. For example, Lennon et al first use a 0.5 ratio and then a 0.7 ratio of beads to hone in on a size range of 300-1000bp.
I would like to try this out for Illumina prep, but I haven't found a lot of details on optimizations for desired size ranges. My goal is a quite narrow size range (400-500bp fragments), and I wonder if the double AMPure approach can do this for me.
Are there any good titration curves out there that would give me an idea of what AMPure ratios would do the job?
Thanks for any insights.
Bjorn
As shown in Lennon et al (2010, Genome Biol.) and Rodrigue et al (2010, Plos One), the Broad and many other folks seem to routinely employ a double AMPure bead approach to obviate the need for gels and to make size selection more scalable. For example, Lennon et al first use a 0.5 ratio and then a 0.7 ratio of beads to hone in on a size range of 300-1000bp.
I would like to try this out for Illumina prep, but I haven't found a lot of details on optimizations for desired size ranges. My goal is a quite narrow size range (400-500bp fragments), and I wonder if the double AMPure approach can do this for me.
Are there any good titration curves out there that would give me an idea of what AMPure ratios would do the job?
Thanks for any insights.
Bjorn