Good day,
Could somebody please elaborate on the ratios of Ampure XP beads required for DOUBLE-SIZE SELECTION to achieve an average fragment size of ~500 bp.
I had followed an illumina protocol: Improved Protocols for Illumina Sequencing (Alternate Protocol 2, Ampure Bead Double DNA Size Selection; https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3849550/).
Briefly, an initial 0.65X addition of reagent:sample was followed by discarding the beads (bound to larger unwanted fragments) and transferring the supernatant to a clean tube. I had then added a 0.12x ratio of reagent to (input) sample volume. The supernatant was then discarded, and the DNA eluted from the beads. However, based on Bio analyzer results, the trace dips at the ~500 bp section but rather produces TWO peaks on either side...
Please help!!!
Could somebody please elaborate on the ratios of Ampure XP beads required for DOUBLE-SIZE SELECTION to achieve an average fragment size of ~500 bp.
I had followed an illumina protocol: Improved Protocols for Illumina Sequencing (Alternate Protocol 2, Ampure Bead Double DNA Size Selection; https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3849550/).
Briefly, an initial 0.65X addition of reagent:sample was followed by discarding the beads (bound to larger unwanted fragments) and transferring the supernatant to a clean tube. I had then added a 0.12x ratio of reagent to (input) sample volume. The supernatant was then discarded, and the DNA eluted from the beads. However, based on Bio analyzer results, the trace dips at the ~500 bp section but rather produces TWO peaks on either side...
Please help!!!
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